Prmt5 inhibitors and uses thereof

ABSTRACT

Described herein are compounds of Formula (I), pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof. Compounds of the present invention are useful for inhibiting PRMT5 activity. Methods of using the compounds for treating PRMT5-mediated disorders are also described.

RELATED APPLICATIONS

This application claims priority under 35 U.S.C. § 119(e) to U.S.provisional patent application, U.S. Ser. No. 61/784,958, filed Mar. 14,2013, and to U.S. provisional patent application, U.S. Ser. No.61/745,393, filed Dec. 21, 2012, the entire contents of each of whichare incorporated herein by reference.

BACKGROUND OF THE INVENTION

Epigenetic regulation of gene expression is an important biologicaldeterminant of protein production and cellular differentiation and playsa significant pathogenic role in a number of human diseases.

Epigenetic regulation involves heritable modification of geneticmaterial without changing its nucleotide sequence. Typically, epigeneticregulation is mediated by selective and reversible modification (e.g.,methylation) of DNA and proteins (e.g., histones) that control theconformational transition between transcriptionally active and inactivestates of chromatin. These covalent modifications can be controlled byenzymes such as methyltransferases (e.g., PRMT5), many of which areassociated with specific genetic alterations that can cause humandisease.

Disease-associated chromatin-modifying enzymes (e.g., PRMT5) play a rolein diseases such as proliferative disorders, metabolic disorders, andblood disorders. Thus, there is a need for the development of smallmolecules that are capable of inhibiting the activity of PRMT5.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

Protein arginine methyltransferase 5 (PRMT5) catalyzes the addition oftwo methyl groups to the two w-guanidino nitrogen atoms of arginine,resulting in ω-NG, N′G symmetric dimethylation of arginine (sDMA) of thetarget protein. PRMT5 functions in the nucleus as well as in thecytoplasm, and its substrates include histones, spliceosomal proteins,transcription factors (See e.g., Sun et al., 2011, PNAS 108:20538-20543). PRMT5 generally functions as part of a molecule weightprotein complex. While the protein complexes of PRMT5 can have a varietyof components, they generally include the protein MEP50 (methylosomeprotein 50). In addition, PRMT5 acts in conjunction with cofactor SAM(S-adenosyl methionine).

PRMT5 is an attractive target for modulation given its role in theregulation of diverse biological processes. It has now been found thatcompounds described herein, and pharmaceutically acceptable salts andcompositions thereof, are effective as inhibitors of PRMT5. Suchcompounds have the general Formula (I):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In some embodiments, pharmaceutical compositions are provided whichcomprise a compound described herein (e.g., a compound of Formula (I)),or a pharmaceutically acceptable salt thereof, and optionally apharmaceutically acceptable excipient.

In certain embodiments, compounds described herein inhibit activity ofPRMT5. In certain embodiments, methods of inhibiting PRMT5 are providedwhich comprise contacting PRMT5 with an effective amount of a compoundof Formula (I), or a pharmaceutically acceptable salt thereof. The PRMT5may be purified or crude, and may be present in a cell, tissue, or asubject. Thus, such methods encompass inhibition of PRMT5 activity bothin vitro and in vivo. In certain embodiments, the PRMT5 is wild-typePRMT5. In certain embodiments, the PRMT5 is overexpressed. In certainembodiments, the PRMT5 is a mutant. In certain embodiments, the PRMT5 isin a cell. In certain embodiments, the PRMT5 is in an animal, e.g., ahuman. In some embodiments, the PRMT5 is in a subject that issusceptible to normal levels of PRMT5 activity due to one or moremutations associated with a PRMT5 substrate. In some embodiments, thePRMT5 is in a subject known or identified as having abnormal PRMT5activity (e.g., overexpression). In some embodiments, a providedcompound is selective for PRMT5 over other methyltransferases. Incertain embodiments, a provided compound is at least about 10-foldselective, at least about 20-fold selective, at least about 30-foldselective, at least about 40-fold selective, at least about 50-foldselective, at least about 60-fold selective, at least about 70-foldselective, at least about 80-fold selective, at least about 90-foldselective, or at least about 100-fold selective relative to one or moreother methyltransferases.

In certain embodiments, methods of altering gene expression in a cellare provided which comprise contacting a cell with an effective amountof a compound of Formula (I), or a pharmaceutically acceptable saltthereof, or a pharmaceutical composition thereof. In certainembodiments, the cell in culture in vitro. In certain embodiments, cellis in an animal, e.g., a human.

In certain embodiments, methods of altering transcription in a cell areprovided which comprise contacting a cell with an effective amount of acompound of Formula (I), or a pharmaceutically acceptable salt thereof,or a pharmaceutical composition thereof. In certain embodiments, thecell in culture in vitro. In certain embodiments, the cell is in ananimal, e.g., a human.

In some embodiments, methods of treating a PRMT5-mediated disorder areprovided which comprise administering to a subject suffering from aPRMT5-mediated disorder an effective amount of a compound describedherein (e.g., a compound of Formula (I)), or a pharmaceuticallyacceptable salt thereof, or a pharmaceutical composition thereof. Incertain embodiments, the PRMT5-mediated disorder is a proliferativedisorder, a metabolic disorder, or a blood disorder. In certainembodiments, compounds described herein are useful for treating cancer.In certain embodiments, compounds described herein are useful fortreating hematopoietic cancer, lung cancer, prostate cancer, melanoma,or pancreatic cancer. In certain embodiments, compounds described hereinare useful for treating a hemoglobinopathy. In certain embodiments,compounds described herein are useful for treating sickle cell anemia.In certain embodiments, compounds described herein are useful fortreating diabetes or obesity.

Compounds described herein are also useful for the study of PRMT5 inbiological and pathological phenomena, the study of intracellular signaltransduction pathways mediated by PRMT5, and the comparative evaluationof new PRMT5 inhibitors.

This application refers to various issued patent, published patentapplications, journal articles, and other publications, all of which areincorporated herein by reference.

Definitions of specific functional groups and chemical terms aredescribed in more detail below. The chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75^(th) Ed., inside cover, andspecific functional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in ThomasSorrell, Organic Chemistry, University Science Books, Sausalito, 1999;Smith and March, March's Advanced Organic Chemistry, 5^(th) Edition,John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive OrganicTransformations, VCH Publishers, Inc., New York, 1989; and Carruthers,Some Modern Methods of Organic Synthesis, 3^(rd) Edition, CambridgeUniversity Press, Cambridge, 1987.

Compounds described herein can comprise one or more asymmetric centers,and thus can exist in various isomeric forms, e.g., enantiomers and/ordiastereomers. For example, the compounds described herein can be in theform of an individual enantiomer, diastereomer or geometric isomer, orcan be in the form of a mixture of stereoisomers, including racemicmixtures and mixtures enriched in one or more stereoisomer. Isomers canbe isolated from mixtures by methods known to those skilled in the art,including chiral high pressure liquid chromatography (HPLC) and theformation and crystallization of chiral salts; or preferred isomers canbe prepared by asymmetric syntheses. See, for example, Jacques et al.,Enantiomers, Racemates and Resolutions (Wiley Interscience, New York,1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistryof Carbon Compounds (McGraw-Hill, N Y, 1962); and Wilen, Tables ofResolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ.of Notre Dame Press, Notre Dame, Ind. 1972). The present disclosureadditionally encompasses compounds described herein as individualisomers substantially free of other isomers, and alternatively, asmixtures of various isomers.

It is to be understood that the compounds of the present invention maybe depicted as different tautomers. It should also be understood thatwhen compounds have tautomeric forms, all tautomeric forms are intendedto be included in the scope of the present invention, and the naming ofany compound described herein does not exclude any tautomer form.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds that differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of hydrogen by deuterium ortritium, replacement of ¹⁹F with ¹⁸F, or the replacement of a carbon bya ¹³C- or ¹⁴C-enriched carbon are within the scope of the disclosure.Such compounds are useful, for example, as analytical tools or probes inbiological assays.

The term “aliphatic,” as used herein, includes both saturated andunsaturated, nonaromatic, straight chain (i.e., unbranched), branched,acyclic, and cyclic (i.e., carbocyclic) hydrocarbons. In someembodiments, an aliphatic group is optionally substituted with one ormore functional groups. As will be appreciated by one of ordinary skillin the art, “aliphatic” is intended herein to include alkyl, alkenyl,alkynyl, cycloalkyl, and cycloalkenyl moieties.

When a range of values is listed, it is intended to encompass each valueand sub-range within the range. For example “C₁₋₆ alkyl” is intended toencompass, C₁, C₂, C₃, C₄, C₅, C₆, C₁₋₆, C₁₋₅, C₁₋₄, C₁₋₃, C₁₋₂, C₂₋₆,C₂₋₅, C₂₋₄, C₂₋₃, C₃₋₆, C₃₋₅, C₃₋₄, C₄₋₆, C₄₋₅, and C₅₋₆ alkyl.

“Alkyl” refers to a radical of a straight-chain or branched saturatedhydrocarbon group having from 1 to 20 carbon atoms (“C₁₋₂₀ alkyl”). Insome embodiments, an alkyl group has 1 to 10 carbon atoms (“C₁₋₁₀alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms(“C₁₋₉ alkyl”). In some embodiments, an alkyl group has 1 to 8 carbonatoms (“C₁₋₈ alkyl”). In some embodiments, an alkyl group has 1 to 7carbon atoms (“C₁₋₇ alkyl”). In some embodiments, an alkyl group has 1to 6 carbon atoms (“C₁₋₆ alkyl”). In some embodiments, an alkyl grouphas 1 to 5 carbon atoms (“C₁₋₅ alkyl”). In some embodiments, an alkylgroup has 1 to 4 carbon atoms (“C₁₋₄ alkyl”). In some embodiments, analkyl group has 1 to 3 carbon atoms (“C₁₋₃ alkyl”). In some embodiments,an alkyl group has 1 to 2 carbon atoms (“C₁₋₂ alkyl”). In someembodiments, an alkyl group has 1 carbon atom (“C₁ alkyl”). In someembodiments, an alkyl group has 2 to 6 carbon atoms (“C₂₋₆ alkyl”).Examples of C₁₋₆ alkyl groups include methyl (C₁), ethyl (C₂), n-propyl(C₃), isopropyl (C₃), n-butyl (C₄), tert-butyl (C₄), sec-butyl (C₄),iso-butyl (C₄), n-pentyl (C₅), 3-pentanyl (C₅), amyl (C₅), neopentyl(C₅), 3-methyl-2-butanyl (C₅), tertiary amyl (C₅), and n-hexyl (C₆).Additional examples of alkyl groups include n-heptyl (C₇), n-octyl (C₈)and the like. In certain embodiments, each instance of an alkyl group isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted alkyl”) or substituted (a “substituted alkyl”) with oneor more substituents. In certain embodiments, the alkyl group isunsubstituted C₁₋₁₀ alkyl (e.g., —CH₃). In certain embodiments, thealkyl group is substituted C₁₋₁₀ alkyl.

In some embodiments, an alkyl group is substituted with one or morehalogens. “Perhaloalkyl” is a substituted alkyl group as defined hereinwherein all of the hydrogen atoms are independently replaced by ahalogen, e.g., fluoro, bromo, chloro, or iodo. In some embodiments, thealkyl moiety has 1 to 8 carbon atoms (“C₁₋₈ perhaloalkyl”). In someembodiments, the alkyl moiety has 1 to 6 carbon atoms (“C₁₋₆perhaloalkyl”). In some embodiments, the alkyl moiety has 1 to 4 carbonatoms (“C₁₋₄ perhaloalkyl”). In some embodiments, the alkyl moiety has 1to 3 carbon atoms (“C₁₋₃ perhaloalkyl”). In some embodiments, the alkylmoiety has 1 to 2 carbon atoms (“C₁₋₂ perhaloalkyl”). In someembodiments, all of the hydrogen atoms are replaced with fluoro. In someembodiments, all of the hydrogen atoms are replaced with chloro.Examples of perhaloalkyl groups include —CF₃, —CF₂CF₃, —CF₂CF₂CF₃,—CCl₃, —CFCl₂, —CF₂Cl, and the like.

“Alkenyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon double bonds, and no triple bonds (“C₂₋₂₀ alkenyl”). Insome embodiments, an alkenyl group has 2 to 10 carbon atoms (“C₂₋₁₀alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms(“C₂₋₉ alkenyl”). In some embodiments, an alkenyl group has 2 to 8carbon atoms (“C₂₋₈ alkenyl”). In some embodiments, an alkenyl group has2 to 7 carbon atoms (“C₂₋₇ alkenyl”). In some embodiments, an alkenylgroup has 2 to 6 carbon atoms (“C₂₋₆ alkenyl”). In some embodiments, analkenyl group has 2 to 5 carbon atoms (“C₂₋₅ alkenyl”). In someembodiments, an alkenyl group has 2 to 4 carbon atoms (“C₂₋₄ alkenyl”).In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C₂₋₃alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C₂alkenyl”). The one or more carbon-carbon double bonds can be internal(such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples ofC₂₋₄ alkenyl groups include ethenyl (C₂), 1-propenyl (C₃), 2-propenyl(C₃), 1-butenyl (C₄), 2-butenyl (C₄), butadienyl (C₄), and the like.Examples of C₂₋₆ alkenyl groups include the aforementioned C₂₋₄ alkenylgroups as well as pentenyl (C₅), pentadienyl (C₅), hexenyl (C₆), and thelike. Additional examples of alkenyl include heptenyl (C₇), octenyl(C₈), octatrienyl (C₈), and the like. In certain embodiments, eachinstance of an alkenyl group is independently optionally substituted,e.g., unsubstituted (an “unsubstituted alkenyl”) or substituted (a“substituted alkenyl”) with one or more substituents. In certainembodiments, the alkenyl group is unsubstituted C₂₋₁₀ alkenyl. Incertain embodiments, the alkenyl group is substituted C₂₋₁₀ alkenyl.

“Alkynyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon triple bonds, and optionally one or more double bonds(“C₂₋₂₀ alkynyl”). In some embodiments, an alkynyl group has 2 to 10carbon atoms (“C₂₋₁₀ alkynyl”). In some embodiments, an alkynyl grouphas 2 to 9 carbon atoms (“C₂₋₉ alkynyl”). In some embodiments, analkynyl group has 2 to 8 carbon atoms (“C₂₋₈ alkynyl”). In someembodiments, an alkynyl group has 2 to 7 carbon atoms (“C₂₋₇ alkynyl”).In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C₂₋₆alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms(“C₂₋₅ alkynyl”). In some embodiments, an alkynyl group has 2 to 4carbon atoms (“C₂₋₄ alkynyl”). In some embodiments, an alkynyl group has2 to 3 carbon atoms (“C₂₋₃ alkynyl”). In some embodiments, an alkynylgroup has 2 carbon atoms (“C₂ alkynyl”). The one or more carbon-carbontriple bonds can be internal (such as in 2-butynyl) or terminal (such asin 1-butynyl). Examples of C₂₋₄ alkynyl groups include, withoutlimitation, ethynyl (C₂), 1-propynyl (C₃), 2-propynyl (C₃), 1-butynyl(C₄), 2-butynyl (C₄), and the like. Examples of C₂₋₆ alkenyl groupsinclude the aforementioned C₂₋₄ alkynyl groups as well as pentynyl (C₅),hexynyl (C₆), and the like. Additional examples of alkynyl includeheptynyl (C₇), octynyl (C₈), and the like. In certain embodiments, eachinstance of an alkynyl group is independently optionally substituted,e.g., unsubstituted (an “unsubstituted alkynyl”) or substituted (a“substituted alkynyl”) with one or more substituents. In certainembodiments, the alkynyl group is unsubstituted C₂₋₁₀ alkynyl. Incertain embodiments, the alkynyl group is substituted C₂₋₁₀ alkynyl.

“Carbocyclyl” or “carbocyclic” refers to a radical of a non-aromaticcyclic hydrocarbon group having from 3 to 10 ring carbon atoms (“C₃₋₁₀carbocyclyl”) and zero heteroatoms in the non-aromatic ring system. Insome embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms(“C₃₋₈ carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to6 ring carbon atoms (“C₃₋₆ carbocyclyl”). In some embodiments, acarbocyclyl group has 3 to 6 ring carbon atoms (“C₃₋₆ carbocyclyl”). Insome embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms(“C₅₋₁₀ carbocyclyl”). Exemplary C₃₋₆ carbocyclyl groups include,without limitation, cyclopropyl (C₃), cyclopropenyl (C₃), cyclobutyl(C₄), cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl (C₅),cyclohexyl (C₆), cyclohexenyl (C₆), cyclohexadienyl (C₆), and the like.Exemplary C₃₋₈ carbocyclyl groups include, without limitation, theaforementioned C₃₋₆ carbocyclyl groups as well as cycloheptyl (C₇),cycloheptenyl (C₇), cycloheptadienyl (C₇), cycloheptatrienyl (C₇),cyclooctyl (C₈), cyclooctenyl (C₈), bicyclo[2.2.1]heptanyl (C₇),bicyclo[2.2.2]octanyl (C₈), and the like. Exemplary C₃₋₁₀ carbocyclylgroups include, without limitation, the aforementioned C₃₋₈ carbocyclylgroups as well as cyclononyl (C₉), cyclononenyl (C₉), cyclodecyl (C₁₀),cyclodecenyl (C₁₀), octahydro-1H-indenyl (C₉), decahydronaphthalenyl(C₁₀), spiro[4.5]decanyl (C₁₀), and the like. As the foregoing examplesillustrate, in certain embodiments, the carbocyclyl group is eithermonocyclic (“monocyclic carbocyclyl”) or contain a fused, bridged orspiro ring system such as a bicyclic system (“bicyclic carbocyclyl”) andcan be saturated or can be partially unsaturated. “Carbocyclyl” alsoincludes ring systems wherein the carbocyclyl ring, as defined above, isfused with one or more aryl or heteroaryl groups wherein the point ofattachment is on the carbocyclyl ring, and in such instances, the numberof carbons continue to designate the number of carbons in thecarbocyclic ring system. In certain embodiments, each instance of acarbocyclyl group is independently optionally substituted, e.g.,unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a“substituted carbocyclyl”) with one or more substituents. In certainembodiments, the carbocyclyl group is unsubstituted C₃₋₁₀ carbocyclyl.In certain embodiments, the carbocyclyl group is a substituted C₃₋₁₀carbocyclyl.

In some embodiments, “carbocyclyl” is a monocyclic, saturatedcarbocyclyl group having from 3 to 10 ring carbon atoms (“C₃₋₁₀cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8 ringcarbon atoms (“C₃₋₈ cycloalkyl”). In some embodiments, a cycloalkylgroup has 3 to 6 ring carbon atoms (“C₃₋₆ cycloalkyl”). In someembodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C₅₋₆cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ringcarbon atoms (“C₅₋₁₀ cycloalkyl”). Examples of C₅₋₆ cycloalkyl groupsinclude cyclopentyl (C₅) and cyclohexyl (C₅). Examples of C₃₋₆cycloalkyl groups include the aforementioned C₅₋₆ cycloalkyl groups aswell as cyclopropyl (C₃) and cyclobutyl (C₄). Examples of C₃₋₈cycloalkyl groups include the aforementioned C₃₋₆ cycloalkyl groups aswell as cycloheptyl (C₇) and cyclooctyl (C₈). In certain embodiments,each instance of a cycloalkyl group is independently unsubstituted (an“unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”)with one or more substituents. In certain embodiments, the cycloalkylgroup is unsubstituted C₃₋₁₀ cycloalkyl. In certain embodiments, thecycloalkyl group is substituted C₃₋₁₀ cycloalkyl.

“Heterocyclyl” or “heterocyclic” refers to a radical of a 3- to10-membered non-aromatic ring system having ring carbon atoms and 1 to 4ring heteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“3-10 membered heterocyclyl”). Inheterocyclyl groups that contain one or more nitrogen atoms, the pointof attachment can be a carbon or nitrogen atom, as valency permits. Aheterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”)or a fused, bridged or spiro ring system such as a bicyclic system(“bicyclic heterocyclyl”), and can be saturated or can be partiallyunsaturated. Heterocyclyl bicyclic ring systems can include one or moreheteroatoms in one or both rings. “Heterocyclyl” also includes ringsystems wherein the heterocyclyl ring, as defined above, is fused withone or more carbocyclyl groups wherein the point of attachment is eitheron the carbocyclyl or heterocyclyl ring, or ring systems wherein theheterocyclyl ring, as defined above, is fused with one or more aryl orheteroaryl groups, wherein the point of attachment is on theheterocyclyl ring, and in such instances, the number of ring memberscontinue to designate the number of ring members in the heterocyclylring system. In certain embodiments, each instance of heterocyclyl isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted heterocyclyl”) or substituted (a “substitutedheterocyclyl”) with one or more substituents. In certain embodiments,the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. Incertain embodiments, the heterocyclyl group is substituted 3-10 memberedheterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 memberednon-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“5-10 membered heterocyclyl”). In someembodiments, a heterocyclyl group is a 5-8 membered non-aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms, wherein eachheteroatom is independently selected from nitrogen, oxygen, and sulfur(“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl groupis a 5-6 membered non-aromatic ring system having ring carbon atoms and1-4 ring heteroatoms, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In someembodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has one ring heteroatomselected from nitrogen, oxygen, and sulfur.

Exemplary 3-membered heterocyclyl groups containing one heteroatominclude, without limitation, azirdinyl, oxiranyl, and thiorenyl.Exemplary 4-membered heterocyclyl groups containing one heteroatominclude, without limitation, azetidinyl, oxetanyl, and thietanyl.Exemplary 5-membered heterocyclyl groups containing one heteroatominclude, without limitation, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl,and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groupscontaining two heteroatoms include, without limitation, dioxolanyl,oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-memberedheterocyclyl groups containing three heteroatoms include, withoutlimitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary6-membered heterocyclyl groups containing one heteroatom include,without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl,and thianyl. Exemplary 6-membered heterocyclyl groups containing twoheteroatoms include, without limitation, piperazinyl, morpholinyl,dithianyl, and dioxanyl. Exemplary 6-membered heterocyclyl groupscontaining two heteroatoms include, without limitation, triazinanyl.Exemplary 7-membered heterocyclyl groups containing one heteroatominclude, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary8-membered heterocyclyl groups containing one heteroatom include,without limitation, azocanyl, oxecanyl, and thiocanyl. Exemplary5-membered heterocyclyl groups fused to a C₆ aryl ring (also referred toherein as a 5,6-bicyclic heterocyclic ring) include, without limitation,indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl,benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclyl groupsfused to an aryl ring (also referred to herein as a 6,6-bicyclicheterocyclic ring) include, without limitation, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and the like.

“Aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclicor tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 πelectrons shared in a cyclic array) having 6-14 ring carbon atoms andzero heteroatoms provided in the aromatic ring system (“C₆₋₁₄ aryl”). Insome embodiments, an aryl group has six ring carbon atoms (“C₆ aryl”;e.g., phenyl). In some embodiments, an aryl group has ten ring carbonatoms (“C₁₀ aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl). Insome embodiments, an aryl group has fourteen ring carbon atoms (“C₁₄aryl”; e.g., anthracyl). “Aryl” also includes ring systems wherein thearyl ring, as defined above, is fused with one or more carbocyclyl orheterocyclyl groups wherein the radical or point of attachment is on thearyl ring, and in such instances, the number of carbon atoms continue todesignate the number of carbon atoms in the aryl ring system. In certainembodiments, each instance of an aryl group is independently optionallysubstituted, e.g., unsubstituted (an “unsubstituted aryl”) orsubstituted (a “substituted aryl”) with one or more substituents. Incertain embodiments, the aryl group is unsubstituted C₆₋₁₄ aryl. Incertain embodiments, the aryl group is substituted C₆₋₁₄ aryl.

“Heteroaryl” refers to a radical of a 5-10 membered monocyclic orbicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electronsshared in a cyclic array) having ring carbon atoms and 1-4 ringheteroatoms provided in the aromatic ring system, wherein eachheteroatom is independently selected from nitrogen, oxygen and sulfur(“5-10 membered heteroaryl”). In heteroaryl groups that contain one ormore nitrogen atoms, the point of attachment can be a carbon or nitrogenatom, as valency permits. Heteroaryl bicyclic ring systems can includeone or more heteroatoms in one or both rings. “Heteroaryl” includes ringsystems wherein the heteroaryl ring, as defined above, is fused with oneor more carbocyclyl or heterocyclyl groups wherein the point ofattachment is on the heteroaryl ring, and in such instances, the numberof ring members continue to designate the number of ring members in theheteroaryl ring system. “Heteroaryl” also includes ring systems whereinthe heteroaryl ring, as defined above, is fused with one or more arylgroups wherein the point of attachment is either on the aryl orheteroaryl ring, and in such instances, the number of ring membersdesignates the number of ring members in the fused (aryl/heteroaryl)ring system. Bicyclic heteroaryl groups wherein one ring does notcontain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and thelike) the point of attachment can be on either ring, e.g., either thering bearing a heteroatom (e.g., 2-indolyl) or the ring that does notcontain a heteroatom (e.g., 5-indolyl).

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-8 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-6 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In someembodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selectedfrom nitrogen, oxygen, and sulfur. In certain embodiments, each instanceof a heteroaryl group is independently optionally substituted, e.g.,unsubstituted (“unsubstituted heteroaryl”) or substituted (“substitutedheteroaryl”) with one or more substituents. In certain embodiments, theheteroaryl group is unsubstituted 5-14 membered heteroaryl. In certainembodiments, the heteroaryl group is substituted 5-14 memberedheteroaryl.

Exemplary 5-membered heteroaryl groups containing one heteroatominclude, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary5-membered heteroaryl groups containing two heteroatoms include, withoutlimitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, andisothiazolyl. Exemplary 5-membered heteroaryl groups containing threeheteroatoms include, without limitation, triazolyl, oxadiazolyl, andthiadiazolyl. Exemplary 5-membered heteroaryl groups containing fourheteroatoms include, without limitation, tetrazolyl. Exemplary6-membered heteroaryl groups containing one heteroatom include, withoutlimitation, pyridinyl. Exemplary 6-membered heteroaryl groups containingtwo heteroatoms include, without limitation, pyridazinyl, pyrimidinyl,and pyrazinyl. Exemplary 6-membered heteroaryl groups containing threeor four heteroatoms include, without limitation, triazinyl andtetrazinyl, respectively. Exemplary 7-membered heteroaryl groupscontaining one heteroatom include, without limitation, azepinyl,oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groupsinclude, without limitation, indolyl, isoindolyl, indazolyl,benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl,benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl,indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groupsinclude, without limitation, naphthyridinyl, pteridinyl, quinolinyl,isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

“Partially unsaturated” refers to a group that includes at least onedouble or triple bond. The term “partially unsaturated” is intended toencompass rings having multiple sites of unsaturation, but is notintended to include aromatic groups (e.g., aryl or heteroaryl groups) asherein defined. Likewise, “saturated” refers to a group that does notcontain a double or triple bond, i.e., contains all single bonds.

In some embodiments, aliphatic, alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aryl, and heteroaryl groups, as defined herein, areoptionally substituted (e.g., “substituted” or “unsubstituted”aliphatic, “substituted” or “unsubstituted” alkyl, “substituted” or“unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl,“substituted” or “unsubstituted” carbocyclyl, “substituted” or“unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or“substituted” or “unsubstituted” heteroaryl group). In general, the term“substituted”, whether preceded by the term “optionally” or not, meansthat at least one hydrogen present on a group (e.g., a carbon ornitrogen atom) is replaced with a permissible substituent, e.g., asubstituent which upon substitution results in a stable compound, e.g.,a compound which does not spontaneously undergo transformation such asby rearrangement, cyclization, elimination, or other reaction. Unlessotherwise indicated, a “substituted” group has a substituent at one ormore substitutable positions of the group, and when more than oneposition in any given structure is substituted, the substituent iseither the same or different at each position. The term “substituted” iscontemplated to include substitution with all permissible substituentsof organic compounds, including any of the substituents described hereinthat results in the formation of a stable compound. The presentdisclosure contemplates any and all such combinations in order to arriveat a stable compound. For purposes of this disclosure, heteroatoms suchas nitrogen may have hydrogen substituents and/or any suitablesubstituent as described herein which satisfy the valencies of theheteroatoms and results in the formation of a stable moiety.

Exemplary carbon atom substituents include, but are not limited to,halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(aa), —ON(R^(bb))₂,—N(R^(bb))₂, —N(R^(bb))₃ ⁺X⁻, —N(OR^(cc))R^(bb), —SH, —SR^(aa),—SSR^(cc), —C(═O)R^(aa), —CO₂H, —CHO, —C(OR^(cc))₂, —CO₂R^(aa),—OC(═O)R^(aa), —OCO₂R^(aa), —C(═O)N(R^(bb))₂, —OC(═O)N(R^(bb))₂,—NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa), —NR^(bb)C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —OC(═NR^(bb))R^(aa),—OC(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂, —OC(═NR^(bb))N(R^(bb))₂,—NR^(bb)C(═NR^(bb))N(R^(bb))₂, —C(═O)NR^(bb)SO₂R^(aa),—NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa), —SO₂OR^(aa), —OSO₂R^(aa),—S(═O)R^(aa), —OS(═O)R^(aa), —Si(R^(aa))₃,—OSi(R^(aa))₃—C(═S)N(R^(bb))₂, —C(═O)SR^(aa), —C(═S)SR^(aa),—SC(═S)SR^(aa), —SC(═O)SR^(aa), —OC(═O)SR^(aa), —SC(═O)OR^(aa),—SC(═O)R^(aa), —P(═O)₂R^(aa), —OP(═O)₂R^(aa), —P(═O)(R^(aa))₂,—OP(═O)(R^(aa))₂, —OP(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂,—OP(═O)₂N(R^(bb))₂, —P(═O)(NR^(bb))₂, —OP(═O)(NR^(bb))₂,—NR^(bb)P(═O)(OR^(cc))₂, —NR^(bb)(═O)(NR^(bb))₂, —P(R^(cc))₂,—P(R^(cc))₃, —OP(R^(cc))₂, —OP(R^(cc))₃, —B(R^(aa))₂, —B(OR^(cc))₂,—BR^(aa)(OR^(cc)), C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

or two geminal hydrogens on a carbon atom are replaced with the group═O, ═S, ═NN(R^(bb))₂, ═NNR^(bb)C(═O)R^(aa), ═NNR^(bb)C(═O)OR^(aa),═NNR^(bb)S(═O)₂R^(aa), ═NR^(bb), or ═NOR^(cc);

each instance of R^(aa) is, independently, selected from C₁₋₁₀ alkyl,C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl,3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl, ortwo R^(aa) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(bb) is, independently, selected from hydrogen, —OH,—OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂,—SO₂R^(cc), —SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc),—C(═S)SR^(cc), —P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂,—P(═O)(NR^(cc))₂, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, or two R^(bb) groups are joined to form a 3-14membered heterocyclyl or 5-14 membered heteroaryl ring, wherein eachalkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroarylis independently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(cc) is, independently, selected from hydrogen, C₁₋₁₀alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups are joined to form a 3-14 memberedheterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl,alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(dd) is, independently, selected from halogen, —CN,—NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(ee), —ON(R^(ff))₂, —N(R^(ff))₂,—N(R^(ff))₃ ⁺X⁻, —N(OR^(ee))R^(ff), —SH, —SR^(ee), —SSR^(ee),—C(═O)R^(ee), —CO₂H, —CO₂R^(ee), —OC(═O)R^(ee), —OCO₂R^(ee),—C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂, —NR^(ff)C(═O)R^(ee),—NR^(ff)CO₂R^(ee), —NR^(ff)C(═O)N(R^(ff))₂, —C(═NR^(ff))OR^(ee),—OC(═NR^(ff))R^(ee), —OC(═NR^(ff))OR^(ee), —C(═NR^(ff))N(R^(ff))₂,—OC(═NR^(ff))N(R^(ff))₂, —NR^(ff)C(═NR^(ff))N(R^(ff))₂,—NR^(ff)SO₂R^(ee), —SO₂N(R^(ff))₂, —SO₂R^(ee), —SO₂OR^(ee), —OSO₂R^(ee),—S(═O)R^(ee), —Si(R^(ee))₃, —OSi(R^(ee))₃, —C(═S)N(R^(ff))₂,—C(═O)SR^(ee), —C(═S)SR^(ee), —SC(═S)SR^(ee), —P(═O)₂R^(ee),—P(═O)(R^(ee))₂, —OP(═O)(R^(ee))₂, —OP(═O)(OR^(ee))₂, C₁₋₆ alkyl, C₁₋₆perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, 3-10membered heterocyclyl, C₆₋₁₀ aryl, 5-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups, or two geminal R^(dd) substituents can be joined to form ═O or═S;

each instance of R^(ee) is, independently, selected from C₁₋₆ alkyl,C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, C₆₋₁₀aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups;

each instance of R^(ff) is, independently, selected from hydrogen, C₁₋₆alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl,3-10 membered heterocyclyl, C₆₋₁₀ aryl and 5-10 membered heteroaryl, ortwo R^(ff) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups; and

each instance of R^(gg) is, independently, halogen, —CN, —NO₂, —N₃,—SO₂H, —SO₃H, —OH, —OC₁₋₆ alkyl, —ON(C₁₋₆ alkyl)₂, —N(C₁₋₆ alkyl)₂,—N(C₁₋₆ alkyl)₃ ⁺X⁻, —NH(C₁₋₆ alkyl)₂ ⁺X⁻, —NH₂(C₁₋₆ alkyl)⁺X⁻, —NH₃⁺X⁻, —N(OC₁₋₆ alkyl)(C₁₋₆ alkyl), —N(OH)(C₁₋₆ alkyl), —NH(OH), —SH,—SC₁₋₆ alkyl, —SS(C₁₋₆ alkyl), —C(═O)(C₁₋₆ alkyl), —CO₂H, —CO₂(C₁₋₆alkyl), —OC(═O)(C₁₋₆ alkyl), —OCO₂(C₁₋₆ alkyl), —C(═O)NH₂, —C(═O)N(C₁₋₆alkyl)₂, —OC(═O)NH(C₁₋₆ alkyl), —NHC(═O)(C₁₋₆ alkyl), —N(C₁₋₆alkyl)C(═O)(C₁₋₆ alkyl), —NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁₋₆ alkyl)₂,—NHC(═O)NH(C₁₋₆ alkyl), —NHC(═O)NH₂, —C(═NH)O(C₁₋₆ alkyl), —OC(═NH)(C₁₋₆alkyl), —OC(═NH)OC₁₋₆ alkyl, —C(═NH)N(C₁₋₆ alkyl)₂, —C(═NH)NH(C₁₋₆alkyl), —C(═NH)NH₂, —OC(═NH)N(C₁₋₆ alkyl)₂, —OC(NH)NH(C₁₋₆ alkyl),—OC(NH)NH₂, —NHC(NH)N(C₁₋₆ alkyl)₂, —NHC(═NH)NH₂, —NHSO₂(C₁₋₆ alkyl),—SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl), —SO₂NH₂, —SO₂C₁₋₆ alkyl,—SO₂OC₁₋₆ alkyl, —OSO₂C₁₋₆ alkyl, —SOC₁₋₆ alkyl, —Si(C₁₋₆ alkyl)₃,—OSi(C₁₋₆ alkyl)₃-C(═S)N(C₁₋₆ alkyl)₂, C(═S)NH(C₁₋₆ alkyl), C(═S)NH₂,—C(═O)S(C₁₋₆ alkyl), —C(═S)SC₁₋₆ alkyl, —SC(═S)SC₁₋₆ alkyl, —P(═O)₂(C₁₋₆alkyl), —P(═O)(C₁₋₆ alkyl)₂, —OP(═O)(C₁₋₆ alkyl)₂, —OP(═O)(OC₁₋₆alkyl)₂, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₃₋₁₀ carbocyclyl, C₆₋₁₀ aryl, 3-10 membered heterocyclyl, 5-10 memberedheteroaryl; or two geminal R^(gg) substituents can be joined to form ═Oor ═S; wherein X⁻ is a counterion.

A “counterion” or “anionic counterion” is a negatively charged groupassociated with a cationic quaternary amino group in order to maintainelectronic neutrality. Exemplary counterions include halide ions (e.g.,F⁻, Cl⁻, Br⁻, I⁻), NO₃ ⁻, ClO₄ ⁻, OH⁻, H₂PO₄ ⁻, HSO₄ ⁻, sulfonate ions(e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate,benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate,naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonicacid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate,ethanoate, propanoate, benzoate, glycerate, lactate, tartrate,glycolate, and the like).

“Halo” or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro,—Cl), bromine (bromo, —Br), or iodine (iodo, —I).

Nitrogen atoms can be substituted or unsubstituted as valency permits,and include primary, secondary, tertiary, and quarternary nitrogenatoms. Exemplary nitrogen atom substitutents include, but are notlimited to, hydrogen, —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —C(═NR^(bb))R^(aa),—C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc),—SO₂OR^(aa), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc),—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂, —P(═O)(NR^(cc))₂,C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups attached to a nitrogen atom are joinedto form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5R^(dd) groups, and wherein R^(aa), R^(bb), R^(cc) and R^(dd) are asdefined above.

In certain embodiments, the substituent present on a nitrogen atom is anitrogen protecting group (also referred to as an amino protectinggroup). Nitrogen protecting groups include, but are not limited to, —OH,—OR^(aa), —N(R^(cc))₂, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))R^(aa), —C(═NR^(cc))OR^(aa),—C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc), —SO₂OR^(cc),—SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc), C₁₋₁₀ alkyl(e.g., aralkyl, heteroaralkyl), C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aralkyl, aryl, and heteroaryl is independently substitutedwith 0, 1, 2, 3, 4, or 5 R^(dd) groups, and wherein R^(aa), R^(bb),R^(cc), and R^(dd) are as defined herein. Nitrogen protecting groups arewell known in the art and include those described in detail inProtecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts,3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference.

Amide nitrogen protecting groups (e.g., —C(═O)R^(aa)) include, but arenot limited to, formamide, acetamide, chloroacetamide,trichloroacetamide, trifluoroacetamide, phenylacetamide,3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide,N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide,o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide,(N′-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide,3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide,2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine,o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

Carbamate nitrogen protecting groups (e.g., —C(═O)OR^(aa)) include, butare not limited to, methyl carbamate, ethyl carbamante,9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate(Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate,1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC),1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC),1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinylcarbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate(Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc),8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate,2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate,2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc),2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate(Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc),1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate,p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate,2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate,3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methylcarbamate, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzylcarbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentylcarbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate,2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate,isobutyl carbamate, isonicotinyl carbamate,p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate,1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate,1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate,p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate,4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzylcarbamate.

Sulfonamide nitrogen protecting groups (e.g., —S(═O)₂R^(aa)) include,but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide,2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),2,4,6-trimethoxybenzenesulfonamide (Mtb),2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide(Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide(Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide,4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

Other nitrogen protecting groups include, but are not limited to,phenothiazinyl-(10)-acyl derivative, N′-p-toluenesulfonylaminoacylderivative, N′-phenylaminothioacyl derivative, N-benzoylphenylalanylderivative, N-acetylmethionine derivative,4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts),N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE),5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine,N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammoniumsalts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),N-9-phenylfluorenylamine (PhF),N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm),N-2-picolylamino N′-oxide, N-1,1-dimethylthiomethyleneamine,N-benzylideneamine, N-p-methoxybenzylideneamine,N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,N—(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine,N-p-nitrobenzylideneamine, N-salicylideneamine,N-5-chlorosalicylideneamine,N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,N-borane derivative, N-diphenylborinic acid derivative,N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate,N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide,diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt),diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzylphosphoramidate, diphenyl phosphoramidate, benzenesulfenamide,o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide,pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

In certain embodiments, the substituent present on an oxygen atom is anoxygen protecting group (also referred to as a hydroxyl protectinggroup). Oxygen protecting groups include, but are not limited to,—R^(aa), —N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, —C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa),—C(═NR^(bb))N(R^(bb))₂, —S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃,—P(R^(cc))₂, —P(R^(cc))₃, —P(═O)₂R^(aa), —P(═O)(R^(aa))₂,—P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and —P(═O)(NR^(bb))₂, whereinR^(aa), R^(bb), and R^(cc) are as defined herein. Oxygen protectinggroups are well known in the art and include those described in detailin Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M.Wuts, 3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference.

Exemplary oxygen protecting groups include, but are not limited to,methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl,(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl,bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR),tetrahydropyranyl (THP), 3-bromotetrahydropyranyl,tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl(MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl,benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl,p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido,diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, o-naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl,1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl(TMS), triethylsilyl (TES), triisopropylsilyl (TIPS),dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS),dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl(TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl,diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate,benzoylformate, acetate, chloroacetate, dichloroacetate,trichloroacetate, trifluoroacetate, methoxyacetate,triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate,3-phenylpropionate, 4-oxopentanoate (levulinate),4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate,adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate,2,4,6-trimethylbenzoate (mesitoate), tert-butyl carbonate (BOC), alkylmethyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethylcarbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc),2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethylcarbonate (Psec), 2-(triphenylphosphonio) ethyl carbonate (Peoc), alkylisobutyl carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkylp-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzylcarbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzylcarbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate,4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate,4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,o-(methoxyacyl)benzoate, o-naphthoate, nitrate, alkylN,N,N′,N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate(Ts).

In certain embodiments, the substituent present on a sulfur atom is asulfur protecting group (also referred to as a thiol protecting group).Sulfur protecting groups include, but are not limited to, —R^(aa),—N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂,—S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃, —P(R^(cc))₂, —P(R^(cc))₃,—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and—P(═O)(NR^(bb))₂, wherein R^(aa), R^(bb) and R^(cc) are as definedherein. Sulfur protecting groups are well known in the art and includethose described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P. G. M. Wuts, 3^(rd) edition, John Wiley & Sons, 1999,incorporated herein by reference.

These and other exemplary substituents are described in more detail inthe Detailed Description, Examples, and claims. The present disclosureis not intended to be limited in any manner by the above exemplarylisting of substituents.

“Pharmaceutically acceptable salt” refers to those salts which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and other animals without undue toxicity,irritation, allergic response, and the like, and are commensurate with areasonable benefit/risk ratio. Pharmaceutically acceptable salts arewell known in the art. For example, Berge et al. describepharmaceutically acceptable salts in detail in J. PharmaceuticalSciences (1977) 66:1-19. Pharmaceutically acceptable salts of thecompounds describe herein include those derived from suitable inorganicand organic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid, or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. Representativealkali or alkaline earth metal salts include sodium, lithium, potassium,calcium, magnesium, and the like. Further pharmaceutically acceptablesalts include, when appropriate, quaternary salts.

A “subject” to which administration is contemplated includes, but is notlimited to, humans (e.g., a male or female of any age group, e.g., apediatric subject (e.g, infant, child, adolescent) or adult subject(e.g., young adult, middle-aged adult or senior adult)) and/or othernon-human animals, for example, non-human mammals (e.g., primates (e.g.,cynomolgus monkeys, rhesus monkeys); commercially relevant mammals suchas cattle, pigs, horses, sheep, goats, cats, and/or dogs), birds (e.g.,commercially relevant birds such as chickens, ducks, geese, and/orturkeys), rodents (e.g., rats and/or mice), reptiles, amphibians, andfish. In certain embodiments, the non-human animal is a mammal. Thenon-human animal may be a male or female at any stage of development. Anon-human animal may be a transgenic animal.

“Condition,” “disease,” and “disorder” are used interchangeably herein.

“Treat,” “treating” and “treatment” encompasses an action that occurswhile a subject is suffering from a condition which reduces the severityof the condition or retards or slows the progression of the condition(“therapeutic treatment”). “Treat,” “treating” and “treatment” alsoencompasses an action that occurs before a subject begins to suffer fromthe condition and which inhibits or reduces the severity of thecondition (“prophylactic treatment”).

An “effective amount” of a compound refers to an amount sufficient toelicit the desired biological response, e.g., treat the condition. Aswill be appreciated by those of ordinary skill in this art, theeffective amount of a compound described herein may vary depending onsuch factors as the desired biological endpoint, the pharmacokinetics ofthe compound, the condition being treated, the mode of administration,and the age and health of the subject. An effective amount encompassestherapeutic and prophylactic treatment.

A “therapeutically effective amount” of a compound is an amountsufficient to provide a therapeutic benefit in the treatment of acondition or to delay or minimize one or more symptoms associated withthe condition. A therapeutically effective amount of a compound means anamount of therapeutic agent, alone or in combination with othertherapies, which provides a therapeutic benefit in the treatment of thecondition. The term “therapeutically effective amount” can encompass anamount that improves overall therapy, reduces or avoids symptoms orcauses of the condition, or enhances the therapeutic efficacy of anothertherapeutic agent.

A “prophylactically effective amount” of a compound is an amountsufficient to prevent a condition, or one or more symptoms associatedwith the condition or prevent its recurrence. A prophylacticallyeffective amount of a compound means an amount of a therapeutic agent,alone or in combination with other agents, which provides a prophylacticbenefit in the prevention of the condition. The term “prophylacticallyeffective amount” can encompass an amount that improves overallprophylaxis or enhances the prophylactic efficacy of anotherprophylactic agent.

As used herein, the term “methyltransferase” represents transferaseclass enzymes that are able to transfer a methyl group from a donormolecule to an acceptor molecule, e.g., an amino acid residue of aprotein or a nucleic base of a DNA molecule. Methytransferases typicallyuse a reactive methyl group bound to sulfur in S-adenosyl methionine(SAM) as the methyl donor. In some embodiments, a methyltransferasedescribed herein is a protein methyltransferase. In some embodiments, amethyltransferase described herein is a histone methyltransferase.Histone methyltransferases (HMT) are histone-modifying enzymes,(including histone-lysine N-methyltransferase and histone-arginineN-methyltransferase), that catalyze the transfer of one or more methylgroups to lysine and arginine residues of histone proteins. In certainembodiments, a methyltransferase described herein is a histone-arginineN-methyltransferase.

As generally described above, provided herein are compounds useful asPRMT5 inhibitors. In some embodiments, the present disclosure provides acompound of Formula (I):

or a pharmaceutically acceptable salt thereof,wherein

represents a single or double bond;

R¹ is hydrogen, R^(z), or —C(O)R^(z), wherein R^(z) is optionallysubstituted C₁₋₆ alkyl;

L is —O—, —N(R)—, —C(R²)(R³)—, —O—CR²R³, —N(R)—CR²R³—, —O—CR²R³—O—,—N(R)—CR²R³—O, —N(R)—CR²R³—N(R)—, —O—CR²R³—N(R)—, —CR²R³—O—,—CR²R³—N(R)—, —O—CR²R³—CR⁹R¹⁰—, —N(R)—CR²R³—CR⁹R¹⁰—, —CR²R³—CR⁹R¹⁰—O—,—CR²R³—CR⁹R¹⁰—N(R)—, or —CR²R³—CR⁹R¹⁰—;

each R is independently hydrogen or optionally substituted C₁₋₆aliphatic;

R² and R³ are independently selected from the group consisting ofhydrogen, halo, —CN, —NO₂, optionally substituted aliphatic, optionallysubstituted carbocyclyl; optionally substituted phenyl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R² andR³ are taken together with their intervening atoms to form an optionallysubstituted carbocyclic or heterocyclic ring; or R² and R³ are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring;

each R^(A) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl;

each R^(B) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl, or two R^(B) groups aretaken together with their intervening atoms to form an optionallysubstituted heterocyclic ring;

Ring A is a monocyclic or bicyclic, saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur;

R⁴ is -L₁-Cy;

L₁ is a bond, —O—, —S—, —N(R)—, —C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—,—N(R)C(O)—, —N(R)C(O)O—, —OC(O)N(R)—, —SO₂—, —SO₂N(R)—, —N(R)SO₂—,—OC(O)—, —C(O)O—, or an optionally substituted, straight or branched,C₁₋₆ aliphatic chain wherein one, two, or three methylene units of L₁are optionally and independently replaced by —O—, —S—, —N(R)—, —C(O)—,—C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)—, —N(R)C(O)O—, —OC(O)N(R)—, —SO₂—,—SO₂N(R)—, —N(R)SO₂—, —OC(O)—, or —C(O)O—;

Cy is an optionally substituted, monocyclic, bicyclic or tricyclic,saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur;

R⁵, R⁶, R⁷, and R⁸ are independently hydrogen, halo, or optionallysubstituted aliphatic;

R⁹ and R¹⁰ are independently selected from the group consisting ofhydrogen, halo, —CN, —NO₂, optionally substituted aliphatic, optionallysubstituted carbocyclyl; optionally substituted phenyl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁹ andR¹⁰ are taken together with their intervening atoms to form anoptionally substituted carbocyclic or heterocyclic ring;

each R^(y) is independently selected from the group consisting of halo,—CN, —NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl; optionally substituted phenyl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂;

each R^(x) is independently selected from the group consisting of halo,—CN, optionally substituted aliphatic, —OR′, and —N(R″)₂;

R′ is hydrogen or optionally substituted aliphatic;

each R″ is independently hydrogen or optionally substituted aliphatic,or two R″ are taken together with their intervening atoms to form aheterocyclic ring;

n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency permits;

m is 0, 1, 2, 3, 4, 5, 6, 7, or 8, as valency permits; and

p is 0 or 1.

In some embodiments, when L is —O— and Ring A is phenyl, p is 1.

In some embodiments, the compound is not one of the following:

In certain embodiments, a provided compound is of Formula (I-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (I-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (I-c):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (I-d):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R⁵, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (I′):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (I′-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (I′-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (I′-c):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (I′-d):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L, R¹,R⁴, R⁵, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (II):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R, R¹,R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (II-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R, R¹,R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (II-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R, R¹,R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (II-c):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R, R¹,R⁴, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (II-d):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R, R¹,R⁴, R⁵, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (III):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R²,R³, R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (III-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R²,R³, R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (III-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R²,R³, R⁴, R⁵, R⁶, R⁷, R⁸, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (III-c):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R²,R³, R⁴, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (III-d):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R²,R³, R⁴, R⁵, R^(y), m, p, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IV):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R⁴,R⁵, R⁶, R⁷, R⁸, R^(y), m, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IV-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R⁴,R⁵, R⁶, R⁷, R⁸, R^(y), m, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IV-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R⁴,R⁵, R⁶, R⁷, R⁸, R^(y), m, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IV-c):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R⁴,R^(y), m, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IV-d):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R⁴,R⁵, R^(y), m, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (V):

or a pharmaceutically acceptable salt thereof, wherein X₁, X₂, X₃, X₄,L, L₁, Cy, R¹, R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (V-a):

or a pharmaceutically acceptable salt thereof, wherein X₁, X₂, X₃, X₄,L, L₁, Cy, R¹, R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (V-b):

or a pharmaceutically acceptable salt thereof, wherein X₁, X₂, X₃, X₄,L, L₁, Cy, R¹, R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (V-c):

or a pharmaceutically acceptable salt thereof, wherein X₁, X₂, X₃, X₄,L, L₁, Cy, R¹, R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (V-d):

or a pharmaceutically acceptable salt thereof, wherein X₁, X₂, X₃, X₄,L, L₁, Cy, R¹, R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VI):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VI-a):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VI-b):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VI-c):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VI-d):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VII):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VII-a):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VII-b):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VII-c):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VII-d):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VIII):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VIII-a):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VIII-b):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VIII-c):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VIII-d):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IX):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IX-a):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IX-b):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IX-c):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IX-d):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (X):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (X-a):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (X-b):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (X-c):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (X-d):

or a pharmaceutically acceptable salt thereof, wherein L, L₁, Cy, R¹,R⁵, R^(x), and n are as defined herein.

In some embodiments,

represents a single bond. In some embodiments,

represents a double bond.

As defined generally above, R¹ is hydrogen, R^(z), or —C(O)R^(z),wherein R^(z) is optionally substituted C₁₋₆ alkyl. In certainembodiments, R¹ is hydrogen. In some embodiments, R¹ is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, R¹ is unsubstituted C₁₋₆alkyl. In certain embodiments, R¹ is methyl, ethyl, or propyl. In someembodiments, R¹ is —C(O)R^(z), wherein R^(z) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, R¹ is —C(O)R^(z), wherein R^(z) isunsubstituted C₁₋₆ alkyl. In certain embodiments, R¹ is acetyl.

As defined generally above, L is —O—, —N(R)—, —C(R²)(R³)—, —O—CR²R³,—N(R)—CR²R³—, —O—CR²R³—O—, —N(R)—CR²R³—O, —N(R)—CR²R³—N(R)—,—O—CR²R³—N(R)—, —CR²R³—O—, —CR²R³—N(R)—, —O—CR²R³—CR⁹R¹⁰—,—N(R)—CR²R³—CR⁹R¹⁰—, —CR²R³—CR⁹R¹⁰—O—, —CR²R³—CR⁹R¹⁰—N(R)—, or—CR²R³—CR⁹R¹⁰—. In certain embodiments, L is —O—, —N(R)—, or —CR²R³—,wherein R, R², and R³ are as described herein. In certain embodiments, Lis —O—. In some embodiments, L is —N(R)—. In certain embodiments, L is—NH—. In certain embodiments, L is —N(R)—, wherein R is optionallysubstituted C₁₋₆ aliphatic. In certain embodiments, L is —N(R)—, whereinR is optionally substituted C₁₋₆ alkyl. In certain embodiments, L is—N(R)—, wherein R is unsubstituted C₁₋₆ alkyl. In certain embodiments, Lis —N(R)—, wherein R is acetyl. In certain embodiments, L is —CH₂—O—. Incertain embodiments, L is —CR²R³—O—. In certain embodiments, L is—CR²R³—N(R)—. In certain embodiments, L is —CH₂—NH—.

As defined generally above, each R is independently hydrogen oroptionally substituted C₁₋₆ aliphatic. In certain embodiments, R ishydrogen. In some embodiments, R is optionally substituted C₁₋₆aliphatic. In some embodiments, R is substituted C₁₋₆ aliphatic. In someembodiments, R is unsubstituted C₁₋₆ aliphatic. In some embodiments, Ris optionally substituted C₁₋₆ alkyl. In some embodiments, R issubstituted C₁₋₆ alkyl. In some embodiments, R is unsubstituted C₁₋₆alkyl. In some embodiments, R is methyl, ethyl, or propyl. In someembodiments, R is substituted with an oxo to give an acyl group.

As defined generally above, R² and R³ are independently selected fromthe group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂. In certain embodiments, R² and R³are independently selected from the group consisting of hydrogen, halo,—CN, —NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted phenyl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—OC(O)R^(A), —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂.

In certain embodiments, R² is hydrogen. In some embodiments, R² is nothydrogen. In some embodiments, R² is halo. In certain embodiments, R² isfluoro. In some embodiments, R² is optionally substituted aliphatic. Incertain embodiments, R² is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R² is optionally substituted C₁₋₆ alkyl. In certainembodiments, R² is substituted C₁₋₆ alkyl. In certain embodiments, R² is—CF₃, CHF₂, or CH₂F. In certain embodiments, R² is unsubstituted C₁₋₆alkyl. In certain embodiments, R² is methyl, ethyl, or propyl. In someembodiments, R² is —CN or —NO₂. In some embodiments, R² is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R² is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R² is—N(R^(B))₂. In certain embodiments, R² is —NHR^(B). In certainembodiments, R² is —NH₂. In certain embodiments, R² is —OR^(A). Incertain embodiments, R² is —OH.

In certain embodiments, R³ is hydrogen. In some embodiments, R³ is nothydrogen. In some embodiments, R³ is halo. In certain embodiments, R³ isfluoro. In some embodiments, R³ is optionally substituted aliphatic. Incertain embodiments, R³ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R³ is optionally substituted C₁₋₆ alkyl. In certainembodiments, R³ is substituted C₁₋₆ alkyl. In certain embodiments, R³ is—CF₃, CHF₂, or CH₂F. In certain embodiments, R³ is unsubstituted C₁₋₆alkyl. In certain embodiments, R³ is methyl, ethyl, or propyl. In someembodiments, R³ is —CN or —NO₂. In some embodiments, R³ is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R³ is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R³ is—N(R^(B))₂. In certain embodiments, R³ is —NHR^(B). In certainembodiments, R³ is —NH₂. In certain embodiments, R³ is —OR^(A). Incertain embodiments, R³ is —OH.

In some embodiments, R² and R³ are the same. In some embodiments, R² andR³ are different. In some embodiments, R² and R³ are each hydrogen. Insome embodiments, R² is hydrogen and R³ is not hydrogen. In someembodiments, R² is hydrogen and R³ is optionally substituted aliphatic.In some embodiments, R² is hydrogen and R³ is C₁₋₆ alkyl. In someembodiments, R² is hydrogen and R³ is methyl. In some embodiments, R² ishydrogen and R³ is ethyl or propyl. In some embodiments, R² is hydrogenand R³ is —CF₃, CHF₂, or CH₂F. In some embodiments, R² is hydrogen andR³ is —N(R^(B))₂ or —OR^(A). In some embodiments, R² is hydrogen and R³is —NH₂. In some embodiments, R² is hydrogen and R³ is —OH. In someembodiments, R² and R³ are not hydrogen. In some embodiments, R² and R³are independently optionally substituted aliphatic. In some embodiments,R² and R³ are methyl. In some embodiments, R² and R³ are taken togetherwith their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring.

As defined generally above, Ring A is a monocyclic or bicyclic,saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, Ring A is aromatic. In certain embodiments, Ring Ais saturated. In certain embodiments, Ring A is monocyclic. In certainembodiments, Ring A is bicyclic.

In certain embodiments, Ring A is phenyl. In certain embodiments, Ring Ais a monocyclic heteroaryl having 1-3 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur. In certain embodiments, Ring A is a5- to 6-membered heteroaryl having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In certain embodiments, RingA is a 5-membered heteroaryl having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur (e.g., furanyl, thienyl,pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyrazolyl,isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl. In certainembodiments, Ring A is a 6-membered heteroaryl having 1-3 nitrogens(e.g., pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, triazinyl). Incertain embodiments, Ring A is pyridyl. In certain embodiments, Ring Ais pyrimidyl. In certain embodiments, Ring A is pyridazinyl. In someembodiments, Ring A is a carbocyclic ring. In some embodiments, Ring Ais a 3- to 8-membered saturated carbocyclic ring. In some embodiments,Ring A is a 3- to 8-membered heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur.

In certain embodiments, Ring A is a bicyclic saturated, partiallyunsaturated, or aromatic ring having 0-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In certain embodiments, RingA is an 8- to 12-membered bicyclic saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, Ring A is an 8- to10-membered bicyclic heteroaryl having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In certain embodiments, RingA is a 9-membered bicyclic heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur (e.g., indolyl,isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl,isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl,benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl,benzisothiazolyl, benzthiadiazolyl, indolizinyl). In certainembodiments, Ring A is a 10-membered bicyclic heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur(e.g., naphthyridinyl, quinolinyl, isoquinolinyl, quinoxalinyl,quinazolinyl. In certain embodiments, Ring A is selected from the groupconsisting of quinoline, benzimidazole, benzopyrazole, quinoxaline,tetrahydroquinoline, tetrahydroisoquinoline, naphthalene,tetrahydronaphthalene, 2,3-dihydrobenzo[b][1,4]dioxine, isoindole,2H-benzo[b][1,4]oxazin-3(4H)-one, 3,4-dihydro-2H-benzo[b][1,4]oxazine,and quinoxalin-2(1H)-one.

As defined generally above, L₁ is a bond, —O—, —S—, —N(R)—, —C(O)—,—C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)—, —N(R)C(O)O—, —OC(O)N(R)—, —SO₂—,—SO₂N(R)—, —N(R)SO₂—, —OC(O)—, —C(O)O—, or an optionally substituted,straight or branched, C₁₋₆ aliphatic chain wherein one, two, or threemethylene units of L₁ are optionally and independently replaced by —O—,—S—, —N(R)—, —C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)—,—N(R)C(O)O—, —OC(O)N(R)—, —SO₂—, —SO₂N(R)—, —N(R)SO₂—, —OC(O)—, or—C(O)O—. In some embodiments, L₁ is a bond. In some embodiments, L₁ is—O—, —S—, or —N(R)—. In some embodiments, L₁ is —C(O)—, —C(O)N(R)—, or—N(R)C(O)—. In some embodiments, L₁ is a C₁₋₆ aliphatic chain whereinone, two, or three methylene units of L₁ are optionally andindependently replaced by —O—, —S—, —N(R)—, —C(O)—, —C(O)N(R)—,—N(R)C(O)N(R)—, —N(R)C(O)—, —N(R)C(O)O—, —OC(O)N(R)—, —SO₂—, —SO₂N(R)—,—N(R)SO₂—, —OC(O)—, or —C(O)O—. In some embodiments, L₁ is a C₁₋₃aliphatic chain wherein one methylene unit of L₁ is optionally replacedby —O—, —S—, —N(R)—, —C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)—,—N(R)C(O)O—, —OC(O)N(R)—, —SO₂—, —SO₂N(R)—, —N(R)SO₂—, —OC(O)—, or—C(O)O—. In some embodiments, L₁ is —CHNH—.

As defined generally above, Cy is an optionally substituted, monocyclic,bicyclic or tricyclic, saturated, partially unsaturated, or aromaticring having 0-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In certain embodiments, Cy is aromatic. In certainembodiments, Cy is saturated. In certain embodiments, Cy is monocyclic.In certain embodiments, Cy is bicyclic. In certain embodiments, Cy istricyclic.

In certain embodiments, Cy is optionally substituted phenyl. In certainembodiments, Cy is an optionally substituted 5- to 6-membered heteroarylhaving 1-3 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, Cy is an optionally substituted5-membered heteroaryl having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur (e.g., furanyl, thienyl, pyrrolyl,oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyrazolyl, isothiazolyl,triazolyl, oxadiazolyl, thiadiazolyl. In certain embodiments, Cy is anoptionally substituted 6-membered heteroaryl having 1-3 nitrogens (e.g.,pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, triazinyl). In certainembodiments, Cy is optionally substituted pyrazole, optionallysubstituted pyridyl, or optionally substituted pyrimidyl. In someembodiments, Cy is an optionally substituted carbocyclic ring. In someembodiments, Cy is an optionally substituted 3- to 8-membered saturatedcarbocyclic ring. In some embodiments, Cy is an optionally substituted3- to 8-membered heterocyclic ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur.

In certain embodiments, Cy is an optionally substituted bicyclicsaturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, Cy is an optionally substituted 8- to 12-memberedbicyclic saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, Cy is an optionally substituted 8- to 10-memberedbicyclic heteroaryl having 1-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, Cy is anoptionally substituted 9- to 10-membered bicyclic heteroaryl having 1-4heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, Cy is an optionally substituted 9-membered bicyclicheteroaryl having 1-3 heteroatoms independently selected from nitrogen,oxygen, and sulfur (e.g., indolyl, isoindolyl, indazolyl,benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl,benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl,indolizinyl). In certain embodiments, Cy is an optionally substituted10-membered bicyclic heteroaryl having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur (e.g., naphthyridinyl,quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl. In certainembodiments, Cy is optionally substituted indazole, optionallysubstituted quinoline, optionally substituted benzimidazole, optionallysubstituted benzothiazole, optionally substituted deazapurine,optionally substituted indole, optionally substituted purine, optionallysubstituted pyrazolopyridine, optionally substituted pyrrolopyridine,optionally substituted pyrroloprimidine, optionally substitutedimidazopyridine, or optionally substituted imidazopyridine.

As defined generally above, R⁵, R⁶, R⁷, and R⁸ are independentlyhydrogen, halo, or optionally substituted aliphatic. In someembodiments, R⁵, R⁶, R⁷, and R⁸ are hydrogen. In some embodiments, R⁶,R⁷, and R⁸ are hydrogen, and R⁵ is optionally substituted aliphatic. Insome embodiments, R⁶, R⁷, and R⁸ are hydrogen, and R⁵ is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R⁶, R⁷, and R⁸ arehydrogen, and R⁵ is optionally substituted C₁₋₃ aliphatic. In someembodiments, R⁶, R⁷, and R⁸ are hydrogen, and R⁵ is methyl. In someembodiments, R⁶, R⁷, and R⁵ are hydrogen, and R⁸ is optionallysubstituted aliphatic. In some embodiments, R⁶, R⁷, and R⁵ are hydrogen,and R⁸ is optionally substituted C₁₋₆ aliphatic. In some embodiments,R⁶, R⁷, and R⁵ are hydrogen, and R⁸ is optionally substituted C₁₋₃aliphatic. In some embodiments, R⁶, R⁷, and R⁵ are hydrogen, and R⁸ ismethyl. In some embodiments, R⁵ is hydrogen. In some embodiments, R⁵ ishalo. In certain embodiments, R⁵ is fluoro. In some embodiments, R⁵ isoptionally substituted C₁₋₆ aliphatic. In some embodiments, R⁵ isoptionally substituted C₁₋₃ alkyl. In certain embodiments, R⁵ is methyl.In some embodiments, R⁶ is hydrogen. In some embodiments, R⁶ is halo. Incertain embodiments, R⁶ is fluoro. In some embodiments, R⁶ is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R⁶ is optionallysubstituted C₁₋₃ alkyl. In certain embodiments, R⁶ is methyl. In someembodiments, R⁷ is hydrogen. In some embodiments, R⁷ is halo. In certainembodiments, R⁷ is fluoro. In some embodiments, R⁷ is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R⁷ is optionallysubstituted C₁₋₃ alkyl. In certain embodiments, R⁷ is methyl. In someembodiments, R⁸ is hydrogen. In some embodiments, R⁸ is halo. In certainembodiments, R⁸ is fluoro. In some embodiments, R⁸ is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R⁸ is optionallysubstituted C₁₋₃ alkyl. In certain embodiments, R⁸ is methyl.

As defined generally above, R⁹ and R¹⁰ are independently selected fromthe group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁹ and R¹⁰ are taken togetherwith their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring. In certain embodiments, R⁹ and R¹⁰ areindependently selected from the group consisting of hydrogen, halo, —CN,—NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted phenyl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—OC(O)R^(A), —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁹ and R¹⁰ are taken togetherwith their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring.

In certain embodiments, R⁹ is hydrogen. In some embodiments, R⁹ is nothydrogen. In some embodiments, R⁹ is halo. In certain embodiments, R⁹ isfluoro. In some embodiments, R⁹ is optionally substituted aliphatic. Incertain embodiments, R⁹ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R⁹ is optionally substituted C₁₋₆ alkyl. In certainembodiments, R⁹ is substituted C₁₋₆ alkyl. In certain embodiments, R⁹ is—CF₃, CHF₂, or CH₂F. In certain embodiments, R⁹ is unsubstituted C₁₋₆alkyl. In certain embodiments, R⁹ is methyl, ethyl, or propyl. In someembodiments, R⁹ is —CN or —NO₂. In some embodiments, R⁹ is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R⁹ is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R⁹ is—N(R^(B))₂. In certain embodiments, R⁹ is —NHR^(B). In certainembodiments, R⁹ is —NH₂. In certain embodiments, R⁹ is —OR^(A). Incertain embodiments, R⁹ is —OH.

In certain embodiments, R¹⁰ is hydrogen. In some embodiments, R¹⁰ is nothydrogen. In some embodiments, R¹⁰ is halo. In certain embodiments, R¹⁰is fluoro. In some embodiments, R¹⁰ is optionally substituted aliphatic.In certain embodiments, R¹⁰ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R¹⁰ is optionally substituted C₁₋₆ alkyl. Incertain embodiments, R¹⁰ is substituted C₁₋₆ alkyl. In certainembodiments, R¹⁰ is —CF₃, CHF₂, or CH₂F. In certain embodiments, R¹⁰ isunsubstituted C₁₋₆ alkyl. In certain embodiments, R¹⁰ is methyl, ethyl,or propyl. In some embodiments, R¹⁰ is —CN or —NO₂. In some embodiments,R¹⁰ is optionally substituted carbocyclyl, optionally substitutedphenyl, optionally substituted heterocyclyl, or optionally substitutedheteroaryl. In some embodiments, R¹⁰ is —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A),—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R¹⁰ is—N(R^(B))₂. In certain embodiments, R¹⁰ is —NHR^(B). In certainembodiments, R¹⁰ is —NH₂. In certain embodiments, R¹⁰ is —OR^(A). Incertain embodiments, R¹⁰ is —OH.

In some embodiments, R⁹ and R¹⁰ are the same. In some embodiments, R⁹and R¹⁰ are different. In some embodiments, R⁹ and R¹⁰ are eachhydrogen. In some embodiments, R⁹ is hydrogen and R¹⁰ is not hydrogen.In some embodiments, R⁹ is hydrogen and R¹⁰ is optionally substitutedaliphatic. In some embodiments, R⁹ is hydrogen and R¹⁰ is C₁₋₆ alkyl. Insome embodiments, R⁹ is hydrogen and R¹⁰ is methyl. In some embodiments,R⁹ is hydrogen and R¹⁰ is ethyl or propyl. In certain embodiments, R⁹and hydrogen and R¹⁰ is —CF₃, CHF₂, or CH₂F. In some embodiments, R⁹ ishydrogen and R¹⁰ is —N(R^(B))₂ or —OR^(A). In some embodiments, R⁹ ishydrogen and R¹⁰ is —NH₂. In some embodiments, R⁹ is hydrogen and R¹⁰ is—OH. In some embodiments, R⁹ and R¹⁰ are not hydrogen. In someembodiments, R⁹ and R¹⁰ are independently optionally substitutedaliphatic. In some embodiments, R⁹ and R¹⁰ are methyl. In someembodiments, R⁹ and R¹⁰ are taken together with their intervening atomsto form an optionally substituted carbocyclic or heterocyclic ring.

As defined generally above, each R^(y) is independently selected fromthe group consisting of halo, —CN, —NO₂, optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedaryl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A),—OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂, wherein R^(A) and R^(B) aredescribed herein. In certain embodiments, each R^(y) is independentlyselected from the group consisting of halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂, wherein R^(A) and R^(B) aredescribed herein.

In some embodiments, at least one R^(y) is halo. In certain embodiments,at least one R^(y) is fluoro. In certain embodiments, at least one R^(y)is chloro. In some embodiments, at least one R^(y) is —CN. In someembodiments, at least one R^(y) is —OR^(A), wherein R^(A) is optionallysubstituted aliphatic. In some embodiments, at least one R^(y) is—OR^(A), wherein R^(A) is unsubstituted C₁₋₆ alkyl. In certainembodiments, at least one R^(y) is methoxy, ethoxy, or propoxy. Incertain embodiments, at least one R^(y) is methoxy. In some embodiments,at least one R^(y) is —OR^(A), wherein R^(A) is substituted C₁₋₆ alkyl.In certain embodiments, at least one R^(y) is —OCH₂CH₂N(CH₃)₂. In someembodiments, at least one R^(y) is —N(R^(B))₂. In some embodiments, atleast one R^(y) is —N(R^(B))₂, wherein each R^(B) is independentlyselected from hydrogen or C₁₋₆ alkyl. In some embodiments, at least oneR^(y) is —NHR^(B). In some embodiments, at least one R^(y) is —N(C₁₋₆alkyl)₂, —NH(C₁₋₆ alkyl), or —NH₂. In certain embodiments, at least oneR^(y) is —NH₂. In certain embodiments, at least one R^(y) is —NHCH₃. Incertain embodiments, at least one R^(y) is —N(CH₃)₂.

In some embodiments, at least one R^(y) is optionally substitutedaliphatic. In certain embodiments, at least one R^(y) is substitutedaliphatic. In certain embodiments, at least one R^(y) is unsubstitutedaliphatic. In some embodiments, at least one R^(y) is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) isunsubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) issubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) ismethyl, ethyl, or propyl. In certain embodiments, at least one R^(y) ismethyl. In certain embodiments, at least one R^(y) is —CF₃, CHF₂, orCH₂F.

In certain embodiments, Cy is selected from the group consisting of:

As defined generally above, each R^(x) is independently selected fromthe group consisting of halo, —CN, optionally substituted aliphatic,—OR′, and —N(R″)₂. In certain embodiments, at least one R^(x) is halo.In certain embodiments, at least one R^(x) is fluoro. In certainembodiments, at least one R^(x) is —CN. In certain embodiments, at leastone R^(x) is optionally substituted aliphatic. In certain embodiments,at least one R^(x) is optionally substituted C₁₋₆ alkyl. In certainembodiments, at least one R^(x) is methyl. In certain embodiments, atleast one R^(x) is —CF₃. In certain embodiments, at least one R^(x) is—OR′ or —N(R″)₂. In certain embodiments, R^(x) is not —OR′ or —N(R″)₂.In certain embodiments, at least one R^(x) is —OCH₃. In certainembodiments, R^(x) is not —OCH₃.

One of ordinary skill in the art will appreciate that an R^(x) group canbe attached anywhere on the tetrahydroisoquinoline ordihydroisoquinoline ring. In certain embodiments, an R^(x) group isattached to the benzene portion of the tetrahydroisoquinoline ordihydroisoquinoline ring. In certain embodiments, an R^(x) group isattached to the tetrahydropyridine or dihydropyridine portion of thetetrahydroisoquinoline or dihydroisoquinoline ring. In certainembodiments, R^(x) groups are attached to both the benzene portion andthe tetrahydropyridine (or dihydropyridine) portion of thetetrahydroisoquinoline (or dihydroisoquinoline) ring. See, for example,the structures shown below:

In certain embodiments, a provided compound is of Formula (XI):

or a pharmaceutically acceptable salt thereof.

As defined generally above, n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, asvalency permits. In certain embodiments, n is 0. In certain embodiments,n is 1. In certain embodiments, n is 2.

In certain embodiments, a provided compound is a compound listed inTable 1, or a pharmaceutically acceptable salt thereof.

TABLE 1 Exemplary Compounds Cmpd LC-MS mz No Structure Exact mass (M +H) 1

380.2464 381.2 2

366.2307 367.2 3

488.2675 489.2 4

394.262 395.0 5

396.2413 397.2 6

378.2056 379.1 7

367.226 368.1 8

365.2355 366.1 9

350.1743 351.1 10

373.2042 374.1 11

398.1397 399.1 12

403.2147 403.9 13

395.2573 395.9 14

380.2464 381.0 15

419.2209 420.2 16

350.163 351.0 17

410.2569 411.2 18

416.177 416.9 19

389.1991 390.0 20

389.1991 390.1 21

380.2464 381.1 22

380.2464 381.1 23

380.2464 381.1 24

394.262 395.1 25

366.2307 367.2 26

391.226 392.0 27

391.226 392.0 28

391.226 391.9 29

410.2569 411.1 30

440.2675 441.0 31

536.2787 537.3 32

394.262 395.2 33

394.262 395.2 34

536.2787 537.2 35

366.2307 367.1 36

378.2671 378.9 37

469.2365 469.9 38

469.2365 469.9 39

379.2624 380.3 40

360.1838 361.2 41

469.2365 469.9 42

469.2365 470.2 43

359.1885 360.1 44

363.1947 364.2 45

413.2103 414.1 46

396.2413 397.0 47

366.2307 367.2 48

349.179 350.1 49

394.262 395.1 50

394.262 395.2 51

394.262 395.2 52

365.2355 366.2 53

393.278 394.3 54

410.2569 411.3 55

451.2147 452.3 56

394.262 395.2 57

363.2198 364.3 58

399.1947 400.2 59

403.1784 404.2 60

413.2103 414.2 61

466.1926 467.2 62

461.1369 462.1 63

437.1661 438.2 64

423.1601 424.2 65

410.1994 411.2 66

420.2525 421.2 67

401.1991 402.2 68

395.1697 396.2 69

393.1496 394.1 70

393.174 394.2 71

389.1991 390.2 72

389.1991 390.2 73

389.1991 390.2 74

389.1991 390.2 75

389.1991 390.2 76

378.1744 379.2 77

377.1791 378.2 78

374.1994 375.2 79

409.2365 410.2 80

427.226 428.2 81

441.2416 442.3 82

519.2522 520.3 83

399.1947 400.2 84

400.1899 401.2 85

417.194 418.2 86

449.1991 450.2 87

427.1759 428.2 88

423.1601 424.1 89

410.1994 411.2 90

403.2147 404.2 91

393.1496 394.2 92

393.1496 394.2 93

384.1838 385.2 94

384.1838 385.2 95

375.1834 376.2 96

375.1834 376.2 97

360.1838 361.2 98

360.1838 361.2 99

423.2522 424.2 100

443.2209 444.2 101

427.226 428.2 102

412.2263 413.2 103

483.2522 484.2 104

475.226 476.2 105

489.2416 490.2 106

413.2103 414.2 107

399.1947 400.1 108

413.2103 414.2 109

415.1896 416.2 110

423.1601 424.2 111

398.1994 399.2 112

377.1791 378.2 113

374.1994 375.2 114

413.2103 414.2 115

413.2103 414.2 116

409.2729 410.2 117

413.2216 414.2 118

413.2216 414.2 119

414.2168 415.2 120

414.2168 415.1 121

414.2168 415.2 122

427.226 428.2 123

416.1558 417.2 124

495.1633 496.2 125

404.2212 405.2 126

472.2362 473.2 127

461.1369 462.1 128

461.1369 462.1 129

452.177 453.2 130

437.1661 438.2 131

437.1661 438.2 132

427.1759 428.2 133

427.1759 428.2 134

423.1601 424.2 135

417.194 418.2 136

418.2005 419.2 137

401.1991 402.2 138

391.1948 392.2 139

378.1744 379.2 140

361.179 362.2 141

414.2056 415.2 142

443.2209 444.2 143

503.2573 504.2 144

499.2471 500.3 145

404.1848 405.2 146

473.2315 474.3 147

412.2151 413.2 148

414.1943 415.2 149

458.2682 459.2 150

445.2617 446.3 151

402.2307 403.3 152

402.1943 403.2 153

391.1896 392.2 154

377.1791 378.2 155

361.179 362.2 156

430.1893 431.2 157

403.1896 404.2 158

393.1689 394.2 159

393.1689 394.2 160

406.2005 407.2 161

404.1848 405.2 162

406.2005 407.2 163

404.1848 405.1 164

404.1848 405.2 165

415.2008 416.2 166

400.1787 401.2 167

451.2147 452.3 168

423.1601 424.2 169

416.21 417.3 170

415.1606 416.2 171

402.1943 403.2 172

384.1838 385.2 173

413.2103 414.3 174

413.2103 414.3 175

352.2151 353.2 176

413.2103 414.3 177

443.2209 444.3 178

519.2522 520.3 179

441.2416 442.2 180

418.1893 419.2 181

457.2365 458.2 182

417.2052 418.2 183

406.2005 407.2 184

404.1848 405.2 185

456.2413 457.2 186

416.21 417.2 187

426.2307 427.3 188

400.1787 401.2 189

414.2056 415.2 190

378.1744 379.2 191

415.2008 416.2 192

427.226 428.2 193

485.2315 486.3 194

480.2525 481.3 195

406.2005 407.2 196

407.1845 408.2 197

420.2161 421.3 198

407.1845 408.3 199

413.2216 414.2 200

477.1319 478.2 201

435.221 436.2 202

470.2318 471.2 203

420.2161 421.2 204

417.2052 418.2 205

414.2168 415.2 206

471.227 472.2 207

414.2168 415.2 208

454.2369 455.2 209

468.2525 469.3 210

414.2056 415.2 211

441.2416 442.2 212

427.226 428.2 213

427.226 428.2 214

447.1714 448.2 215

447.1714 448.1 216

418.2369 419.2 217

413.2216 414.2 218

468.2525 469.3 219

482.2682 483.2 220

482.2682 483.3 221

496.2838 497.3 222

527.2533 528.2 223

399.1947 400.1 224

414.2168 415.2 225

427.1896 428.2 226

413.2216 414.2 227

412.2263 413.2 228

524.2787 525.3 229

454.2481 455.2 230

468.2638 469.2 231

511.2583 512.2 232

525.274 526.2 233

511.2696 512.2 234

527.2645 528.2 235

398.2107 399.2 236

413.2216 414.2 237

413.2216 414.2 238

414.2168 415.2 239

414.2168 415.2 240

468.2638 469.2 241

482.2794 483.3 242

524.29 525.4 243

525.274 526.2 244

540.2849 541.2 245

471.2383 472.2 246

525.2852 526.2 247

540.2961 541.2 248

414.2056 415.2 249

524.2787 525.3 250

482.2794 483.3 251

483.2634 484.3 252

496.2951 497.3 253

483.2634 484.3 254

455.2434 456.2 255

483.2747 484.3 256

525.2852 526.3 257

410.1994 411.1 258

410.1994 411.1 259

406.2005 407.2 260

406.2005 407.2 261

413.2216 414.2 262

470.2318 471.3 263

510.2631 511.3 264

526.258 527.3 265

455.2321 456.2 266

413.2103 414.2 267

403.1896 404.2 268

403.1896 404.2 269

455.2209 456.2 270

539.2896 540.4 271

402.2168 403.2 272

402.2168 403.2 273

454.2369 455.2 274

392.1848 393.2 275

413.2216 414.3 276

413.2216 414.3 277

449.1915 450.1 278

331.1339 332.1 279

377.1991 378.0 280

365.0949 366.0 281

365.0949 366.0 282

298.1681 299.1 283

298.1681 299.1 284

298.1681 299.1 285

327.1471 327.9 286

382.262 383.2 287

376.2362 377.2 288

327.1834 328.0 289

318.2307 319.1 290

362.2206 363.1 291

390.2519 390.9 292

390.2519 391.2 293

304.2151 305.1 294

303.2198 304.2 295

315.1947 316.0 296

277.1678 278.1 297

290.1994 291.1 298

290.1994 291.1 299

312.1838 313.0 300

355.1784 356.1 301

323.1885 324.0 302

275.1885 276.1 303

261.1729 262.1 304

305.1991 306.1 305

291.1834 292.1 306

276.1838 277.1 307

290.1994 291.1 308

297.1729 297.9 309

303.2198 304.1 310

291.1834 292.1

In certain embodiments, a provided compound inhibits PRMT5. In certainembodiments, a provided compound inhibits wild-type PRMT5. In certainembodiments, a provided compound inhibits a mutant PRMT5. In certainembodiments, a provided compound inhibits PRMT5, e.g., as measured in anassay described herein. In certain embodiments, the PRMT5 is from ahuman. In certain embodiments, a provided compound inhibits PRMT5 at anIC₅₀ less than or equal to 10 μM. In certain embodiments, a providedcompound inhibits PRMT5 at an IC₅₀ less than or equal to 1 μM. Incertain embodiments, a provided compound inhibits PRMT5 at an IC₅₀ lessthan or equal to 0.1 μM. In certain embodiments, a provided compoundinhibits PRMT5 in a cell at an EC₅₀ less than or equal to 10 μM. Incertain embodiments, a provided compound inhibits PRMT5 in a cell at anEC₅₀ less than or equal to 1 μM. In certain embodiments, a providedcompound inhibits PRMT5 in a cell at an EC₅₀ less than or equal to 0.1μM. In certain embodiments, a provided compound inhibits cellproliferation at an EC₅₀ less than or equal to 10 μM. In certainembodiments, a provided compound inhibits cell proliferation at an EC₅₀less than or equal to 1 μM. In certain embodiments, a provided compoundinhibits cell proliferation at an EC₅₀ less than or equal to 0.1 μM. Insome embodiments, a provided compound is selective for PRMT5 over othermethyltransferases. In certain embodiments, a provided compound is atleast about 10-fold selective, at least about 20-fold selective, atleast about 30-fold selective, at least about 40-fold selective, atleast about 50-fold selective, at least about 60-fold selective, atleast about 70-fold selective, at least about 80-fold selective, atleast about 90-fold selective, or at least about 100-fold selective forPRMT5 relative to one or more other methyltransferases.

It will be understood by one of ordinary skill in the art that the PRMT5can be wild-type PRMT5, or any mutant or variant of PRMT5.

In certain embodiments, the PRMT5 is isoform A (GenBank accession no.NP006100) (SEQ ID NO.:1):

MAAMAVGGAG GSRVSSGRDL NCVPEIADTL GAVAKQGFDFLCMPVFHPRF KREFIQEPAK NRPGPQTRSD LLLSGRDWNTLIVGKLSPWI RPDSKVEKIR RNSEAAMLQE LNFGAYLGLPAFLLPLNQED NTNLARVLTN HIHTGHHSSM FWMRVPLVAPEDLRDDIIEN APTTHTEEYS GEEKTWMWWH NFRTLCDYSKRIAVALEIGA DLPSNHVIDR WLGEPIKAAI LPTSIFLTNKKGFPVLSKMH QRLIFRLLKL EVQFIITGTN HHSEKEFCSYLQYLEYLSQN RPPPNAYELF AKGYEDYLQS PLQPLMDNLESQTYEVFEKD PIKYSQYQQA IYKCLLDRVP EEEKDTNVQVLMVLGAGRGP LVNASLRAAK QADRRIKLYA VEKNPNAVVTLENWQFEEWG SQVTVVSSDM REWVAPEKAD IIVSELLGSFADNELSPECL DGAQHFLKDD GVSIPGEYTS FLAPISSSKLYNEVRACREK DRDPEAQFEM PYVVRLHNFH QLSAPQPCFTFSHPNRDPMI DNNRYCTLEF PVEVNTVLHG FAGYFETVLYQDITLSIRPE THSPGMFSWF PILFPIKQPI TVREGQTICVRFWRCSNSKK VWYEWAVTAP VCSAIHNPTG RSYTIGL

In certain embodiments, the PRMT5 is isoform B (GenBank accession no.NP001034708) (SEQ ID NO.:2)

MRGPNSGTEK GRLVIPEKQG FDFLCMPVFH PRFKREFIQEPAKNRPGPQT RSDLLLSGRD WNTLIVGKLS PWIRPDSKVEKIRRNSEAAM LQELNFGAYL GLPAFLLPLN QEDNTNLARVLTNHIHTGHH SSMFWMRVPL VAPEDLRDDI IENAPTTHTEEYSGEEKTWM WWHNFRTLCD YSKRIAVALE IGADLPSNHVIDRWLGEPIK AAILPTSIFL TNKKGFPVLS KMHQRLIFRLLKLEVQFIIT GTNHHSEKEF CSYLQYLEYL SQNRPPPNAYELFAKGYEDY LQSPLQPLMD NLESQTYEVF EKDPIKYSQYQQAIYKCLLD RVPEEEKDTN VQVLMVLGAG RGPLVNASLRAAKQADRRIK LYAVEKNPNA VVTLENWQFE EWGSQVTVVSSDMREWVAPE KADIIVSELL GSFADNELSP ECLDGAQHFLKDDGVSIPGE YTSFLAPISS SKLYNEVRAC REKDRDPEAQFEMPYVVRLH NFHQLSAPQP CFTFSHPNRD PMIDNNRYCTLEFPVEVNTV LHGFAGYFET VLYQDITLSI RPETHSPGMFSWFPILFPIK QPITVREGQT ICVRFWRCSN SKKVWYEWAV TAPVCSAIHN PTGRSYTIGL

In certain embodiments, the PRMT5 is transcript variant 1 (GenBankaccession no. NM_006109).

The present disclosure provides pharmaceutical compositions comprising acompound described herein, e.g., a compound of Formula (I), or apharmaceutically acceptable salt thereof, as described herein, andoptionally a pharmaceutically acceptable excipient. It will beunderstood by one of ordinary skill in the art that the compoundsdescribed herein, or salts thereof, may be present in various forms,such as hydrates, solvates, or polymorphs. In certain embodiments, aprovided composition comprises two or more compounds described herein.In certain embodiments, a compound described herein, or apharmaceutically acceptable salt thereof, is provided in an effectiveamount in the pharmaceutical composition. In certain embodiments, theeffective amount is a therapeutically effective amount. In certainembodiments, the effective amount is an amount effective for inhibitingPRMT5. In certain embodiments, the effective amount is an amounteffective for treating a PRMT5-mediated disorder. In certainembodiments, the effective amount is a prophylactically effectiveamount. In certain embodiments, the effective amount is an amounteffective to prevent a PRMT5-mediated disorder.

Pharmaceutically acceptable excipients include any and all solvents,diluents, or other liquid vehicles, dispersions, suspension aids,surface active agents, isotonic agents, thickening or emulsifyingagents, preservatives, solid binders, lubricants, and the like, assuited to the particular dosage form desired. General considerations informulation and/or manufacture of pharmaceutical compositions agents canbe found, for example, in Remington's Pharmaceutical Sciences, SixteenthEdition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980), andRemington: The Science and Practice of Pharmacy, 21st Edition(Lippincott Williams & Wilkins, 2005).

Pharmaceutical compositions described herein can be prepared by anymethod known in the art of pharmacology. In general, such preparatorymethods include the steps of bringing a compound described herein (the“active ingredient”) into association with a carrier and/or one or moreother accessory ingredients, and then, if necessary and/or desirable,shaping and/or packaging the product into a desired single- ormulti-dose unit.

Pharmaceutical compositions can be prepared, packaged, and/or sold inbulk, as a single unit dose, and/or as a plurality of single unit doses.As used herein, a “unit dose” is discrete amount of the pharmaceuticalcomposition comprising a predetermined amount of the active ingredient.The amount of the active ingredient is generally equal to the dosage ofthe active ingredient which would be administered to a subject and/or aconvenient fraction of such a dosage such as, for example, one-half orone-third of such a dosage.

Relative amounts of the active ingredient, the pharmaceuticallyacceptable excipient, and/or any additional ingredients in apharmaceutical composition of the present disclosure will vary,depending upon the identity, size, and/or condition of the subjecttreated and further depending upon the route by which the composition isto be administered. By way of example, the composition may comprisebetween 0.1% and 100% (w/w) active ingredient.

Pharmaceutically acceptable excipients used in the manufacture ofprovided pharmaceutical compositions include inert diluents, dispersingand/or granulating agents, surface active agents and/or emulsifiers,disintegrating agents, binding agents, preservatives, buffering agents,lubricating agents, and/or oils. Excipients such as cocoa butter andsuppository waxes, coloring agents, coating agents, sweetening,flavoring, and perfuming agents may also be present in the composition.

Exemplary diluents include calcium carbonate, sodium carbonate, calciumphosphate, dicalcium phosphate, calcium sulfate, calcium hydrogenphosphate, sodium phosphate lactose, sucrose, cellulose,microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodiumchloride, dry starch, cornstarch, powdered sugar, and mixtures thereof.

Exemplary granulating and/or dispersing agents include potato starch,corn starch, tapioca starch, sodium starch glycolate, clays, alginicacid, guar gum, citrus pulp, agar, bentonite, cellulose and woodproducts, natural sponge, cation-exchange resins, calcium carbonate,silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone)(crospovidone), sodium carboxymethyl starch (sodium starch glycolate),carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose(croscarmellose), methylcellulose, pregelatinized starch (starch 1500),microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate,quaternary ammonium compounds, and mixtures thereof.

Exemplary surface active agents and/or emulsifiers include naturalemulsifiers (e.g., acacia, agar, alginic acid, sodium alginate,tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk,casein, wool fat, cholesterol, wax, and lecithin), colloidal clays(e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminumsilicate)), long chain amino acid derivatives, high molecular weightalcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetinmonostearate, ethylene glycol distearate, glyceryl monostearate, andpropylene glycol monostearate, polyvinyl alcohol), carbomers (e.g.,carboxy polymethylene, polyacrylic acid, acrylic acid polymer, andcarboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g.,carboxymethylcellulose sodium, powdered cellulose, hydroxymethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylenesorbitan monolaurate (Tween 20), polyoxyethylene sorbitan (Tween 60),polyoxyethylene sorbitan monooleate (Tween 80), sorbitan monopalmitate(Span 40), sorbitan monostearate (Span 60], sorbitan tristearate (Span65), glyceryl monooleate, sorbitan monooleate (Span 80)),polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj 45),polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil,polyoxymethylene stearate, and Solutol), sucrose fatty acid esters,polyethylene glycol fatty acid esters (e.g., Cremophor™),polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether (Brij 30)),poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamineoleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyllaurate, sodium lauryl sulfate, Pluronic F68, Poloxamer 188, cetrimoniumbromide, cetylpyridinium chloride, benzalkonium chloride, docusatesodium, and/or mixtures thereof.

Exemplary binding agents include starch (e.g., cornstarch and starchpaste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin,molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums(e.g., acacia, sodium alginate, extract of Irish moss, panwar gum,ghatti gum, mucilage of isapol husks, carboxymethylcellulose,methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose,cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate(Veegum), and larch arabogalactan), alginates, polyethylene oxide,polyethylene glycol, inorganic calcium salts, silicic acid,polymethacrylates, waxes, water, alcohol, and/or mixtures thereof.

Exemplary preservatives include antioxidants, chelating agents,antimicrobial preservatives, antifungal preservatives, alcoholpreservatives, acidic preservatives, and other preservatives.

Exemplary antioxidants include alpha tocopherol, ascorbic acid, acorbylpalmitate, butylated hydroxyanisole, butylated hydroxytoluene,monothioglycerol, potassium metabisulfite, propionic acid, propylgallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, andsodium sulfite.

Exemplary chelating agents include ethylenediaminetetraacetic acid(EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodiumedetate, trisodium edetate, calcium disodium edetate, dipotassiumedetate, and the like), citric acid and salts and hydrates thereof(e.g., citric acid monohydrate), fumaric acid and salts and hydratesthereof, malic acid and salts and hydrates thereof, phosphoric acid andsalts and hydrates thereof, and tartaric acid and salts and hydratesthereof. Exemplary antimicrobial preservatives include benzalkoniumchloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide,cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol,chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea,phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate,propylene glycol, and thimerosal.

Exemplary antifungal preservatives include butyl paraben, methylparaben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoicacid, potassium benzoate, potassium sorbate, sodium benzoate, sodiumpropionate, and sorbic acid.

Exemplary alcohol preservatives include ethanol, polyethylene glycol,phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate,and phenylethyl alcohol.

Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E,beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbicacid, sorbic acid, and phytic acid. Other preservatives includetocopherol, tocopherol acetate, deteroxime mesylate, cetrimide,butylated hydroxyanisol (BHA), butylated hydroxytoluened (BHT),ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ethersulfate (SLES), sodium bisulfite, sodium metabisulfite, potassiumsulfite, potassium metabisulfite, Glydant Plus, Phenonip, methylparaben,Germall 115, Germaben II, Neolone, Kathon, and Euxyl. In certainembodiments, the preservative is an anti-oxidant. In other embodiments,the preservative is a chelating agent.

Exemplary buffering agents include citrate buffer solutions, acetatebuffer solutions, phosphate buffer solutions, ammonium chloride, calciumcarbonate, calcium chloride, calcium citrate, calcium glubionate,calcium gluceptate, calcium gluconate, D-gluconic acid, calciumglycerophosphate, calcium lactate, propanoic acid, calcium levulinate,pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasiccalcium phosphate, calcium hydroxide phosphate, potassium acetate,potassium chloride, potassium gluconate, potassium mixtures, dibasicpotassium phosphate, monobasic potassium phosphate, potassium phosphatemixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodiumcitrate, sodium lactate, dibasic sodium phosphate, monobasic sodiumphosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide,aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline,Ringer's solution, ethyl alcohol, and mixtures thereof.

Exemplary lubricating agents include magnesium stearate, calciumstearate, stearic acid, silica, talc, malt, glyceryl behanate,hydrogenated vegetable oils, polyethylene glycol, sodium benzoate,sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate,sodium lauryl sulfate, and mixtures thereof.

Exemplary natural oils include almond, apricot kernel, avocado, babassu,bergamot, black current seed, borage, cade, camomile, canola, caraway,carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee,corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed,geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate,jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademianut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange,orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed,pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood,sasquana, savoury, sea buckthorn, sesame, shea butter, silicone,soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, andwheat germ oils. Exemplary synthetic oils include, but are not limitedto, butyl stearate, caprylic triglyceride, capric triglyceride,cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate,mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixturesthereof.

Liquid dosage forms for oral and parenteral administration includepharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active ingredients,the liquid dosage forms may comprise inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed,groundnut, corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can include adjuvants such as wetting agents, emulsifyingand suspending agents, sweetening, flavoring, and perfuming agents. Incertain embodiments for parenteral administration, the compoundsdescribed herein are mixed with solubilizing agents such as Cremophor™,alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins,polymers, and mixtures thereof.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation can be a sterile injectable solution,suspension or emulsion in a nontoxic parenterally acceptable diluent orsolvent, for example, as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that can be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables. Theinjectable formulations can be sterilized, for example, by filtrationthrough a bacterial-retaining filter, or by incorporating sterilizingagents in the form of sterile solid compositions which can be dissolvedor dispersed in sterile water or other sterile injectable medium priorto use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This can be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Compositions for rectal or vaginal administration are typicallysuppositories which can be prepared by mixing the compounds describedherein with suitable non-irritating excipients or carriers such as cocoabutter, polyethylene glycol or a suppository wax which are solid atambient temperature but liquid at body temperature and therefore melt inthe rectum or vaginal cavity and release the active ingredient.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activeingredient is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may comprise buffering agents.

Solid compositions of a similar type can be employed as fillers in softand hard-filled gelatin capsules using such excipients as lactose ormilk sugar as well as high molecular weight polyethylene glycols and thelike. The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes. Solid compositions of asimilar type can be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active ingredient can be in micro-encapsulated form with one or moreexcipients as noted above. The solid dosage forms of tablets, dragees,capsules, pills, and granules can be prepared with coatings and shellssuch as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active ingredient can be admixed with at least oneinert diluent such as sucrose, lactose, or starch. Such dosage forms maycomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets, and pills, the dosage forms may comprise bufferingagents. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes.

Dosage forms for topical and/or transdermal administration of a providedcompound may include ointments, pastes, creams, lotions, gels, powders,solutions, sprays, inhalants and/or patches. Generally, the activeingredient is admixed under sterile conditions with a pharmaceuticallyacceptable carrier and/or any desired preservatives and/or buffers ascan be required. Additionally, the present disclosure encompasses theuse of transdermal patches, which often have the added advantage ofproviding controlled delivery of an active ingredient to the body. Suchdosage forms can be prepared, for example, by dissolving and/ordispensing the active ingredient in the proper medium. Alternatively oradditionally, the rate can be controlled by either providing a ratecontrolling membrane and/or by dispersing the active ingredient in apolymer matrix and/or gel.

Suitable devices for use in delivering intradermal pharmaceuticalcompositions described herein include short needle devices such as thosedescribed in U.S. Pat. Nos. 4,886,499; 5,190,521; 5,328,483; 5,527,288;4,270,537; 5,015,235; 5,141,496; and 5,417,662. Intradermal compositionscan be administered by devices which limit the effective penetrationlength of a needle into the skin, such as those described in PCTpublication WO 99/34850 and functional equivalents thereof. Jetinjection devices which deliver liquid vaccines to the dermis via aliquid jet injector and/or via a needle which pierces the stratumcorneum and produces a jet which reaches the dermis are suitable. Jetinjection devices are described, for example, in U.S. Pat. Nos.5,480,381; 5,599,302; 5,334,144; 5,993,412; 5,649,912; 5,569,189;5,704,911; 5,383,851; 5,893,397; 5,466,220; 5,339,163; 5,312,335;5,503,627; 5,064,413; 5,520,639; 4,596,556; 4,790,824; 4,941,880;4,940,460; and PCT publications WO 97/37705 and WO 97/13537. Ballisticpowder/particle delivery devices which use compressed gas to acceleratevaccine in powder form through the outer layers of the skin to thedermis are suitable. Alternatively or additionally, conventionalsyringes can be used in the classical mantoux method of intradermaladministration.

Formulations suitable for topical administration include, but are notlimited to, liquid and/or semi liquid preparations such as liniments,lotions, oil in water and/or water in oil emulsions such as creams,ointments and/or pastes, and/or solutions and/or suspensions.Topically-administrable formulations may, for example, comprise fromabout 1% to about 10% (w/w) active ingredient, although theconcentration of the active ingredient can be as high as the solubilitylimit of the active ingredient in the solvent. Formulations for topicaladministration may further comprise one or more of the additionalingredients described herein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation suitable for pulmonary administration via thebuccal cavity. Such a formulation may comprise dry particles whichcomprise the active ingredient and which have a diameter in the rangefrom about 0.5 to about 7 nanometers or from about 1 to about 6nanometers. Such compositions are conveniently in the form of drypowders for administration using a device comprising a dry powderreservoir to which a stream of propellant can be directed to dispersethe powder and/or using a self propelling solvent/powder dispensingcontainer such as a device comprising the active ingredient dissolvedand/or suspended in a low-boiling propellant in a sealed container. Suchpowders comprise particles wherein at least 98% of the particles byweight have a diameter greater than 0.5 nanometers and at least 95% ofthe particles by number have a diameter less than 7 nanometers.Alternatively, at least 95% of the particles by weight have a diametergreater than 1 nanometer and at least 90% of the particles by numberhave a diameter less than 6 nanometers. Dry powder compositions mayinclude a solid fine powder diluent such as sugar and are convenientlyprovided in a unit dose form.

Low boiling propellants generally include liquid propellants having aboiling point of below 65° F. at atmospheric pressure. Generally thepropellant may constitute 50 to 99.9% (w/w) of the composition, and theactive ingredient may constitute 0.1 to 20% (w/w) of the composition.The propellant may further comprise additional ingredients such as aliquid non-ionic and/or solid anionic surfactant and/or a solid diluent(which may have a particle size of the same order as particlescomprising the active ingredient).

Pharmaceutical compositions formulated for pulmonary delivery mayprovide the active ingredient in the form of droplets of a solutionand/or suspension. Such formulations can be prepared, packaged, and/orsold as aqueous and/or dilute alcoholic solutions and/or suspensions,optionally sterile, comprising the active ingredient, and mayconveniently be administered using any nebulization and/or atomizationdevice. Such formulations may further comprise one or more additionalingredients including, but not limited to, a flavoring agent such assaccharin sodium, a volatile oil, a buffering agent, a surface activeagent, and/or a preservative such as methylhydroxybenzoate. The dropletsprovided by this route of administration may have an average diameter inthe range from about 0.1 to about 200 nanometers.

Formulations described herein as being useful for pulmonary delivery areuseful for intranasal delivery of a pharmaceutical composition. Anotherformulation suitable for intranasal administration is a coarse powdercomprising the active ingredient and having an average particle fromabout 0.2 to 500 micrometers. Such a formulation is administered byrapid inhalation through the nasal passage from a container of thepowder held close to the nares.

Formulations for nasal administration may, for example, comprise fromabout as little as 0.1% (w/w) and as much as 100% (w/w) of the activeingredient, and may comprise one or more of the additional ingredientsdescribed herein. A provided pharmaceutical composition can be prepared,packaged, and/or sold in a formulation for buccal administration. Suchformulations may, for example, be in the form of tablets and/or lozengesmade using conventional methods, and may contain, for example, 0.1 to20% (w/w) active ingredient, the balance comprising an orallydissolvable and/or degradable composition and, optionally, one or moreof the additional ingredients described herein. Alternately,formulations for buccal administration may comprise a powder and/or anaerosolized and/or atomized solution and/or suspension comprising theactive ingredient. Such powdered, aerosolized, and/or aerosolizedformulations, when dispersed, may have an average particle and/ordroplet size in the range from about 0.1 to about 200 nanometers, andmay further comprise one or more of the additional ingredients describedherein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation for ophthalmic administration. Such formulationsmay, for example, be in the form of eye drops including, for example, a0.1/1.0% (w/w) solution and/or suspension of the active ingredient in anaqueous or oily liquid carrier. Such drops may further comprisebuffering agents, salts, and/or one or more other of the additionalingredients described herein. Other opthalmically-administrableformulations which are useful include those which comprise the activeingredient in microcrystalline form and/or in a liposomal preparation.Ear drops and/or eye drops are contemplated as being within the scope ofthis disclosure.

Although the descriptions of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions which aresuitable for administration to humans, it will be understood by theskilled artisan that such compositions are generally suitable foradministration to animals of all sorts. Modification of pharmaceuticalcompositions suitable for administration to humans in order to renderthe compositions suitable for administration to various animals is wellunderstood, and the ordinarily skilled veterinary pharmacologist candesign and/or perform such modification with ordinary experimentation.

Compounds provided herein are typically formulated in dosage unit formfor ease of administration and uniformity of dosage. It will beunderstood, however, that the total daily usage of provided compositionswill be decided by the attending physician within the scope of soundmedical judgment. The specific therapeutically effective dose level forany particular subject or organism will depend upon a variety of factorsincluding the disease, disorder, or condition being treated and theseverity of the disorder; the activity of the specific active ingredientemployed; the specific composition employed; the age, body weight,general health, sex and diet of the subject; the time of administration,route of administration, and rate of excretion of the specific activeingredient employed; the duration of the treatment; drugs used incombination or coincidental with the specific active ingredientemployed; and like factors well known in the medical arts.

The compounds and compositions provided herein can be administered byany route, including enteral (e.g., oral), parenteral, intravenous,intramuscular, intra-arterial, intramedullary, intrathecal,subcutaneous, intraventricular, transdermal, interdermal, rectal,intravaginal, intraperitoneal, topical (as by powders, ointments,creams, and/or drops), mucosal, nasal, bucal, sublingual; byintratracheal instillation, bronchial instillation, and/or inhalation;and/or as an oral spray, nasal spray, and/or aerosol. Specificallycontemplated routes are oral administration, intravenous administration(e.g., systemic intravenous injection), regional administration viablood and/or lymph supply, and/or direct administration to an affectedsite. In general the most appropriate route of administration willdepend upon a variety of factors including the nature of the agent(e.g., its stability in the environment of the gastrointestinal tract),and/or the condition of the subject (e.g., whether the subject is ableto tolerate oral administration).

The exact amount of a compound required to achieve an effective amountwill vary from subject to subject, depending, for example, on species,age, and general condition of a subject, severity of the side effects ordisorder, identity of the particular compound(s), mode ofadministration, and the like. The desired dosage can be delivered threetimes a day, two times a day, once a day, every other day, every thirdday, every week, every two weeks, every three weeks, or every fourweeks. In certain embodiments, the desired dosage can be delivered usingmultiple administrations (e.g., two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen, fourteen, or moreadministrations).

In certain embodiments, an effective amount of a compound foradministration one or more times a day to a 70 kg adult human maycomprise about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg,about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosageform.

In certain embodiments, a compound described herein may be administeredat dosage levels sufficient to deliver from about 0.001 mg/kg to about1000 mg/kg, from about 0.01 mg/kg to about mg/kg, from about 0.1 mg/kgto about 40 mg/kg, from about 0.5 mg/kg to about 30 mg/kg, from about0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, orfrom about 1 mg/kg to about 25 mg/kg, of subject body weight per day,one or more times a day, to obtain the desired therapeutic effect.

In some embodiments, a compound described herein is administered one ormore times per day, for multiple days. In some embodiments, the dosingregimen is continued for days, weeks, months, or years.

It will be appreciated that dose ranges as described herein provideguidance for the administration of provided pharmaceutical compositionsto an adult. The amount to be administered to, for example, a child oran adolescent can be determined by a medical practitioner or personskilled in the art and can be lower or the same as that administered toan adult.

It will be also appreciated that a compound or composition, as describedherein, can be administered in combination with one or more additionaltherapeutically active agents. In certain embodiments, a compound orcomposition provided herein is administered in combination with one ormore additional therapeutically active agents that improve itsbioavailability, reduce and/or modify its metabolism, inhibit itsexcretion, and/or modify its distribution within the body. It will alsobe appreciated that the therapy employed may achieve a desired effectfor the same disorder, and/or it may achieve different effects.

The compound or composition can be administered concurrently with, priorto, or subsequent to, one or more additional therapeutically activeagents. In certain embodiments, the additional therapeutically activeagent is a compound of Formula (I). In certain embodiments, theadditional therapeutically active agent is not a compound of Formula(I). In general, each agent will be administered at a dose and/or on atime schedule determined for that agent. In will further be appreciatedthat the additional therapeutically active agent utilized in thiscombination can be administered together in a single composition oradministered separately in different compositions. The particularcombination to employ in a regimen will take into account compatibilityof a provided compound with the additional therapeutically active agentand/or the desired therapeutic effect to be achieved. In general, it isexpected that additional therapeutically active agents utilized incombination be utilized at levels that do not exceed the levels at whichthey are utilized individually. In some embodiments, the levels utilizedin combination will be lower than those utilized individually.

Exemplary additional therapeutically active agents include, but are notlimited to, small organic molecules such as drug compounds (e.g.,compounds approved by the U.S. Food and Drug Administration as providedin the Code of Federal Regulations (CFR)), peptides, proteins,carbohydrates, monosaccharides, oligosaccharides, polysaccharides,nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides orproteins, small molecules linked to proteins, glycoproteins, steroids,nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides,antisense oligonucleotides, lipids, hormones, vitamins, and cells.

Also encompassed by the present disclosure are kits (e.g.,pharmaceutical packs). The kits provided may comprise a providedpharmaceutical composition or compound and a container (e.g., a vial,ampule, bottle, syringe, and/or dispenser package, or other suitablecontainer). In some embodiments, provided kits may optionally furtherinclude a second container comprising a pharmaceutical excipient fordilution or suspension of a provided pharmaceutical composition orcompound. In some embodiments, a provided pharmaceutical composition orcompound provided in the container and the second container are combinedto form one unit dosage form. In some embodiments, a provided kitsfurther includes instructions for use.

Compounds and compositions described herein are generally useful for theinhibition of PRMT5. In some embodiments, methods of treatingPRMT5-mediated disorder in a subject are provided which compriseadministering an effective amount of a compound described herein (e.g.,a compound of Formula (I)), or a pharmaceutically acceptable saltthereof), to a subject in need of treatment. In certain embodiments, theeffective amount is a therapeutically effective amount. In certainembodiments, the effective amount is a prophylactically effectiveamount. In certain embodiments, the subject is suffering from aPRMT5-mediated disorder. In certain embodiments, the subject issusceptible to a PRMT5-mediated disorder.

As used herein, the term “PRMT5-mediated disorder” means any disease,disorder, or other pathological condition in which PRMT5 is known toplay a role. Accordingly, in some embodiments, the present disclosurerelates to treating or lessening the severity of one or more diseases inwhich PRMT5 is known to play a role.

In some embodiments, the present disclosure provides a method ofinhibiting PRMT5 comprising contacting PRMT5 with an effective amount ofa compound described herein (e.g., a compound of Formula (I)), or apharmaceutically acceptable salt thereof. The PRMT5 may be purified orcrude, and may be present in a cell, tissue, or subject. Thus, suchmethods encompass both inhibition of in vitro and in vivo PRMT5activity. In certain embodiments, the method is an in vitro method,e.g., such as an assay method. It will be understood by one of ordinaryskill in the art that inhibition of PRMT5 does not necessarily requirethat all of the PRMT5 be occupied by an inhibitor at once. Exemplarylevels of inhibition of PRMT5 include at least 10% inhibition, about 10%to about 25% inhibition, about 25% to about 50% inhibition, about 50% toabout 75% inhibition, at least 50% inhibition, at least 75% inhibition,about 80% inhibition, about 90% inhibition, and greater than 90%inhibition.

In some embodiments, provided is a method of inhibiting PRMT5 activityin a subject in need thereof comprising administering to the subject aneffective amount of a compound described herein (e.g., a compound ofFormula (I)), or a pharmaceutically acceptable salt thereof.

In certain embodiments, provided is a method of altering gene expressionin a cell which comprises contacting a cell with an effective amount ofa compound of Formula (I), or a pharmaceutically acceptable saltthereof. In certain embodiments, the cell in culture in vitro. Incertain embodiments, the cell is in an animal, e.g., a human. In certainembodiments, the cell is in a subject in need of treatment.

In certain embodiments, provided is a method of altering transcriptionin a cell which comprises contacting a cell with an effective amount ofa compound of Formula (I), or a pharmaceutically acceptable saltthereof. In certain embodiments, the cell in culture in vitro. Incertain embodiments, the cell is in an animal, e.g., a human. In certainembodiments, the cell is in a subject in need of treatment.

In certain embodiments, a method is provided of selecting a therapy fora subject having a disease associated with PRMT5-mediated disorder ormutation comprising the steps of determining the presence ofPRMT5-mediated disorder or gene mutation in the PRMT5 gene or andselecting, based on the presence of PRMT5-mediated disorder a genemutation in the PRMT5 gene a therapy that includes the administration ofa provided compound. In certain embodiments, the disease is cancer.

In certain embodiments, a method of treatment is provided for a subjectin need thereof comprising the steps of determining the presence ofPRMT5-mediated disorder or a gene mutation in the PRMT5 gene andtreating the subject in need thereof, based on the presence of aPRMT5-mediated disorder or gene mutation in the PRMT5 gene with atherapy that includes the administration of a provided compound. Incertain embodiments, the subject is a cancer patient.

In some embodiments, a provided compound is useful in treating aproliferative disorder, such as cancer, a benign neoplasm, an autoimmunedisease, or an inflammatory disease. For example, while not being boundto any particular mechanism, PRMT5 has been shown to be involved incyclin D1 dysregulated cancers. Increased PRMT5 activity mediates keyevents associated with cyclin D1-dependent neoplastic growth includingCUL4 repression, CDT1 overexpression, and DNA re-replication. Further,human cancers harboring mutations in Fbx4, the cyclin D1 E3 ligase,exhibit nuclear cyclin D1 accumulation and increased PRMT5 activity(Aggarwal et al., Cancer Cell. 2010 18(4):329-40). Additionally, PRMT5has also been implicated in accelerating cell cycle progression throughG1 phase and modulating regulators of G1; for example, PRMT5 mayupregulate cyclin-dependent kinase (CDK) 4, CDK6, and cyclins D1, D2 andE1. Moreover, PRMT5 may activate phosphoinositide 3-kinase (PI3K)/AKTsignaling (Wei et al., Cancer Sci. 2012 103(9):1640-50). Thus in someembodiments, the inhibition of PRMT5 by a provided compound is useful intreating the following non-limiting list of cancers: breast cancer,esophageal cancer, bladder cancer, lung cancer, hematopoietic cancer,lymphoma, medulloblastoma, rectum adenocarcinoma, colon adenocarcinoma,gastric cancer, pancreatic cancer, liver cancer, adenoid cysticcarcinoma, lung adenocarcinoma, head and neck squamous cell carcinoma,brain tumors, hepatocellular carcinoma, renal cell carcinoma, melanoma,oligodendroglioma, ovarian clear cell carcinoma, and ovarian serouscystadenocarcinoma.

In some embodiments, the inhibition of PRMT5 by a provided compound isuseful in treating prostate cancer and lung cancer, in which PRMT5 hasbeen shown to play a role (Gu et al., PLoS One 2012; 7(8):e44033; Gu etal., Biochem. J. (2012) 446 (235-241)). In some embodiments, a providedcompound is useful to delay the onset of, slow the progression of, orameliorate the symptoms of cancer. In some embodiments, a providedcompound is administered in combination with other compounds, drugs, ortherapeutics to treat cancer. In some embodiments, compounds describedherein are useful for treating a cancer including, but not limited to,acoustic neuroma, adenocarcinoma, adrenal gland cancer, anal cancer,angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma,hemangiosarcoma), appendix cancer, benign monoclonal gammopathy, biliarycancer (e.g., cholangiocarcinoma), bladder cancer, breast cancer (e.g.,adenocarcinoma of the breast, papillary carcinoma of the breast, mammarycancer, medullary carcinoma of the breast), brain cancer (e.g.,meningioma; glioma, e.g., astrocytoma, oligodendroglioma;medulloblastoma), bronchus cancer, carcinoid tumor, cervical cancer(e.g., cervical adenocarcinoma), choriocarcinoma, chordoma,craniopharyngioma, colorectal cancer (e.g., colon cancer, rectal cancer,colorectal adenocarcinoma), epithelial carcinoma, ependymoma,endotheliosarcoma (e.g., Kaposi's sarcoma, multiple idiopathichemorrhagic sarcoma), endometrial cancer (e.g., uterine cancer, uterinesarcoma), esophageal cancer (e.g., adenocarcinoma of the esophagus,Barrett's adenocarinoma), Ewing sarcoma, eye cancer (e.g., intraocularmelanoma, retinoblastoma), familiar hypereosinophilia, gall bladdercancer, gastric cancer (e.g., stomach adenocarcinoma), gastrointestinalstromal tumor (GIST), head and neck cancer (e.g., head and neck squamouscell carcinoma, oral cancer (e.g., oral squamous cell carcinoma (OSCC),throat cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngealcancer, oropharyngeal cancer)), hematopoietic cancers (e.g., leukemiasuch as acute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL),acute myelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronicmyelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chroniclymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL); lymphoma suchas Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkinlymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma(DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)), follicularlymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma(CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas(e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodalmarginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma),primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacyticlymphoma (i.e., “Waldenström's macroglobulinemia”), hairy cell leukemia(HCL), immunoblastic large cell lymphoma, precursor B-lymphoblasticlymphoma and primary central nervous system (CNS) lymphoma; and T-cellNHL such as precursor T-lymphoblastic lymphomalleukemia, peripheralT-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g.,mycosis fungiodes, Sezary syndrome), angioimmunoblastic T-cell lymphoma,extranodal natural killer T-cell lymphoma, enteropathy type T-celllymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplasticlarge cell lymphoma); a mixture of one or more leukemiallymphoma asdescribed above; and multiple myeloma (MM)), heavy chain disease (e.g.,alpha chain disease, gamma chain disease, mu chain disease),hemangioblastoma, inflammatory myofibroblastic tumors, immunocyticamyloidosis, kidney cancer (e.g., nephroblastoma a.k.a. Wilms' tumor,renal cell carcinoma), liver cancer (e.g., hepatocellular cancer (HCC),malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma, smallcell lung cancer (SCLC), non-small cell lung cancer (NSCLC),adenocarcinoma of the lung), leiomyosarcoma (LMS), mastocytosis (e.g.,systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma,myeloproliferative disorder (MPD) (e.g., polycythemia Vera (PV),essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a.myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocyticleukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilicsyndrome (HES)), neuroblastoma, neurofibroma (e.g., neurofibromatosis(NF) type 1 or type 2, schwannomatosis), neuroendocrine cancer (e.g.,gastroenteropancreatic neuroendoctrine tumor (GEP-NET), carcinoidtumor), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarianembryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma,pancreatic cancer (e.g., pancreatic andenocarcinoma, intraductalpapillary mucinous neoplasm (IPMN), Islet cell tumors), penile cancer(e.g., Paget's disease of the penis and scrotum), pinealoma, primitiveneuroectodermal tumor (PNT), prostate cancer (e.g., prostateadenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary gland cancer,skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA),melanoma, basal cell carcinoma (BCC)), small bowel cancer (e.g.,appendix cancer), soft tissue sarcoma (e.g., malignant fibroushistiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor(MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous glandcarcinoma, sweat gland carcinoma, synovioma, testicular cancer (e.g.,seminoma, testicular embryonal carcinoma), thyroid cancer (e.g.,papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC),medullary thyroid cancer), urethral cancer, vaginal cancer and vulvarcancer (e.g., Paget's disease of the vulva).

In some embodiments, a provided compound is useful in treating ametabolic disorder, such as diabetes or obesity. For example, while notbeing bound to any particular mechanism, a role for PRMT5 has beenrecognized in adipogenesis. Inhibition of PRMT5 expression in multiplecell culture models for adipogenesis prevented the activation ofadipogenic genes, while overexpression of PRMT5 enhanced adipogenic geneexpression and differentiation (LeBlanc et al., Mol Endocrinol. 2012April; 26(4):583-97). Additionally, it has been shown that adipogenesisplays a pivotal role in the etiology and progression of diabetes andobesity (Camp et al., Trends Mol Med. 2002 September; 8(9):442-7). Thusin some embodiments, the inhibition of PRMT5 by a provided compound isuseful in treating diabetes and/or obesity.

In some embodiments, a provided compound is useful to delay the onsetof, slow the progression of, or ameliorate the symptoms of, diabetes. Insome embodiments, the diabetes is Type 1 diabetes. In some embodiments,the diabetes is Type 2 diabetes. In some embodiments, a providedcompound is useful to delay the onset of, slow the progression of, orameliorate the symptoms of, obesity. In some embodiments, a providedcompound is useful to make a subject lose weight. In some embodiments, aprovided compound could be used in combination with other compounds,drugs, or therapeutics, such as metformin and insulin, to treat diabetesand/or obesity.

In some embodiments, a provided compound is useful in treating a blooddisorder, e.g., a hemoglobinopathy, such as sickle cell disease orβ-thalassemia. For example, while not being bound to any particularmechanism, PRMT5 is a known repressor of γ-globin gene expression, andincreased fetal γ-globin (HbF) levels in adulthood are associated withsymptomatic amelioration in sickle cell disease and β-thalassemia (Xu etal., Haematologica. 2012 November; 97(11):1632-40). Thus in someembodiments, the inhibition of PRMT5 by a provided compound is useful intreating a blood disorder, such as a hemoglobinopathy such as sicklecell disease or β-thalassemia.

In some embodiments, a provided compound is useful to delay the onsetof, slow the progression of, or ameliorate the symptoms of, sickle celldisease. In some embodiments, a provided compound is useful to delay theonset of, slow the progression of, or ameliorate the symptoms of,β-thalassemia. In some embodiments, a provided compound could be used incombination with other compounds, drugs, or therapeutics, to treat ahemoglobinopathy such as sickle cell disease or β-thalassemia.

In some embodiments, compounds described herein can prepared usingmethods shown in general Scheme 1, comprising a ring opening of a chiralor racemic epoxide group.

In some embodiments, the epoxide opening is the final step in thesynthesis, as shown in exemplary Scheme 2.

In some embodiments, epoxide opening is employed to build keyintermediates for addition synthesis as shown in exemplary schemes 3-6.

Examples

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

Synthetic Methods Compound 51-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(((tetrahydro-2H-pyran-4-yl)amino)methyl)phenoxy)propan-2-ol

Step 1: 3-(oxiran-2-ylmethoxy)benzaldehyde

To a solution of 3-hydroxybenzaldehyde (2.0 g, 16.38 mmol) in DMF (25mL) was added NaH (60%, 982.6 mg, 24.57 mmol) at 0° C. in portions. Themixture was stirred at this temperature for 30 minutes before a solutionof 2-(chloromethyl)oxirane (2.27 g, 24.57 mmol) in DMF (5 mL) was slowlyadded into the reaction mixture which was then allowed to reach roomtemperature slowly and stirred for 16 h. The solvent was removed byconcentration and the residue dissolved in ethyl acetate and washed withwater. T the separated organic layer was dried over sodium sulfate,filtered and concentrated. The crude product was then purified by columnchromatography. (2.2 g, yield 75%) MS (ESI⁺) e/z: 179.1 [M+1]⁺.

Step 2:3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxropoxy)benzaldehyde

A solution of 3-(oxiran-2-ylmethoxy)benzaldehyde (1.0 g, 5.61 mmol) and1,2,3,4-tetrahydroisoquinoline (1.49 g, 11.22 mmol) in EtOH (20 mL) washeated at 110° C. for 3 h. The solvent was then removed by concentrationand the residue dissolved in ethyl acetate, washed with water and theseparated organic layer dried and concentrated. The crude product waspurified by column chromatography. (1.1 g, yield 63%.) MS (ESI⁺) e/z:312.3 [M+1]⁺.

Step 3:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(((tetrahydro-2H-pyran-4-yl)amino)methyl)phenoxy)propan-2-ol

To a solution of3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)benzaldehyde(200 mg, 0.64 mmol) in DCM (15 mL) was added tetrahydro-2H-pyran-4-amine(97 mg, 0.96 mmol). The solution was heated at 60° C. for 0.5 h. Then,NaBH(OAc)₃ (204 mg, 0.96 mmol) was added into the reaction mixture andheated at 60° C. for another 1 h. The reaction mixture was quenched byadding 1 N HCl aqueous solution and diluted with DCM then washed withNaHCO₃ aqueous solution. The separated organic layer was concentratedand the crude product was purified by preparative HPLC purification. (40mg, yield 17%) MS (ESI⁺) e/z: 397.2 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δppm: 7.36 (t, J=8.0 Hz, 1H), 7.19-7.15 (m, 3H), 7.11-7.02 (m, 4H),4.39-4.33 (m, 1H), 4.16 (s, 2H), 4.08-4.00 (m, 6H), 3.49-3.40 (m, 3H),3.25-3.20 (m, 2H), 3.15-3.03 (m, 4H), 2.12-2.02 (m, 2H), 1.71-1.60 (m,2H).

Compound 61-(3-(((1H-pyrazol-3-yl)amino)methyl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)benzaldehyde(200 mg, 0.64 mmol) in DCM (15 mL) was added 1H-pyrazol-3-amine (80 mg,0.96 mmol). The solution was heated at 60° C. for 0.5 h. NaBH(OAc)₃ (204mg, 0.96 mmol) was then added into the reaction mixture which was heatedat 60° C. for another 1 h. The reaction mixture was quenched by adding 1N HCl aqueous solution and diluted with DCM and washed with NaHCO₃aqueous solution. The separated organic layer was concentrated and thecrude product was purified by preparative HPLC purification. (26 mg,yield 11%). MS (ESI⁺) e/z: 379.1 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm:7.33-7.18 (m, 7H), 7.00-6.96 (m, 2H), 6.84-6.80 (m, 1H), 4.48-4.44 (m,3H), 4.28 (s, 1H), 4.06-3.98 (m, 2H), 3.59-3.56 (m, 2H), 3.42-3.39 (m,2H), 3.20-3.15 (m, 2H).

Compound 31(4-((5-(4-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)-1H-pyrazol-1-yl)methyl)phenyl)(pyrrolidin-1-yl)methanone

Step 1: methyl 4-((5-bromo-1H-pyrazol-1-yl)methyl)benzoate

5-bromo-1H-pyrazole (146 mg, 1 mmol), methyl 4-(bromomethyl)benzoate(242 mg, 2 mmol) and K₂C0₃ (276 mg, 2 mmol) were placed in a 100-mLflask with 2-butanone (20 mL) and heated at reflux temperature for 17 h.The mixture was cooled, filtered and evaporated in vacuo leaving aresidue which was dissolved in ethyl acetate and washed with water. Theseparated organic layer was dried and concentrated to give desiredproduct as an oil (188 mg, 100% yield). This crude material was usedwithout further purification. LCMS (m/z): 295.1 [M+H]⁺

Step 2: 4-((5-bromo-1H-pyrazol-1-yl)methyl)benzoic acid

Methyl 4-((5-bromo-1H-pyrazol-1-yl)methyl)benzoate (264 mg, 0.95 mmol),NaOH (200 mg, 5 mmol) were placed in a 100-mL flask with covered withMeOH and H₂O (1:1) and stirred for 4 h at room temperature. The solutionwas then evaporated, taken up in water and acidified with 2M HCl withthe resulting the solid filtered off yielding the desired product as awhite solid (200 mg, 90%). LCMS (m/z): 281.1 [M+H]⁺

Step 3:(4-((5-bromo-1H-pyrazol-1-yl)methyl)phenyl)(pyrrolidin-1-yl)methanone

4-((5-bromo-1H-pyrazol-1-yl)methyl)benzoic acid (200 mg, 0.71 mmol),pyrrolidine (61 mg, 0.85 mmol) and EDC-HOBt (452 mg, 2 mmol) werecombined in CH₂Cl₂ and stirred for 4 h at room temperature. The mixturewas evaporated in vacuo and the residue dissolved in ethyl acetate andwashed with water. The separated organic layer was dried andconcentrated affording the product as an oil (231 mg, yield 99%), whichwas used in next step without further purification. LCMS (m/z): 334.2[M+H]⁺

Step 4:(4-((5-(4-hydroxyphenyl)-1H-pyrazol-1-yl)methyl)phenyl)(pyrrolidin-1-yl)methanone

To a solution of(4-((5-bromo-1H-pyrazol-1-yl)methyl)phenyl)(pyrrolidin-1-yl) methanone(231 mg, 0.7 mmol) in dioxane:water (5:1, 25 ml) was added(4-hydroxyphenyl)boronic acid (207 mg, 1.5 mmol), Cs₂CO₃ (487 mg, 1.5mmol) and Pd(dppf)Cl₂ (86 mg) and the mixture purged with nitrogen. Themixture was then stirred at 140° C. under microwave mediated heatingirradiation for 40 min. The mixture was diluted with water, extractedwith ethyl acetate three times and the organic layer dried over Na₂SO₄before concentration. The crude product was then purified by columnchromatography to yield the desired product (173 mg, yield 70%). LCMS(m/z): 348.2 [M+H]⁺

Step 5:(4-((5-(4-(oxiran-2-ylmethoxy)phenyl)-1H-pyrazol-1-yl)methyl)phenyl)(pyrrolidin-1-yl)methanone

(4-((5-(4-hydroxyphenyl)-1H-pyrazol-1-yl)methyl)phenyl)(pyrrolidin-1-yl)methanone(173 mg, 0.5 mmol), 2-(bromomethyl)oxirane (544 mg, 3 mmol) and K₂CO₃(552 mg, 4 mmol) were charged to a 100-mL flask containing 2-butanone(20 mL) and heated at reflux temperature for 17 h. The mixture wascooled, filtered and evaporated in vacuo to give crude product as an oil(201 mg, 98%). This crude material was used without furtherpurification. LCMS (m/z): 404.2 [M+H]⁺

Step 6:(4-((5-(4-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)-1H-pyrazol-1-yl)methyl)phenyl)(pyrrolidin-1-yl)methanone

(4-((5-(4-(oxiran-2-ylmethoxy)phenyl)-1H-pyrazol-1-yl)methyl)phenyl)(pyrrolidin-1-yl)methanone(201 mg, 0.5 mmol) was dissolved in absolute ethanol (5 mL) with1,2,3,4-tetrahydroisoquinoline (133 mg, 1 mmol) and heated to 130° C.under microwave heating for 25 min. The sample was concentrated in vacuoand the residue purified by preparative HPLC to give desired titleproduct as the formate salt (120 mg, yield 45%).

¹H NMR (400 MHz, METHANOL-d₄) δ=7.62 (d, J=1.6 Hz, 1H), 7.44 (d, J=8.0Hz, 2H), 7.33-7.20 (m, 6H), 7.06-7.03 (m, 4H), 6.41 (d, J=1.6 Hz, 1H),5.44 (s, 2H), 4.51-4.48 (m, 3H), 4.09 (d, J=4.4 Hz, 2H), 3.64-3.55 (m,4H), 3.43-3.40 (m, 4H), 3.25-3.20 (m, 2H), 2.00-1.87 (m, 4H). LCMS(m/z): 537.3 [M+H]⁺

Compound 391-((3-((cyclopentylamino)methyl)phenyl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

Step 1: 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline

To a stirred solution of 1,2,3,4-tetrahydroisoquinoline (2.0 g, 7.52mmol) in 25 mL CH₃CN was added K₂CO₃ (1.25 g, 9.02 mmol) and thesolution stirred at room temperature for 5 min. 2-(Bromomethyl)oxirane(1.03 g, 7.52 mmol) in MeCN (25 mL) was added drop wise over a 20 minperiod and the reaction mixture stirred at room temperature for 12 h.The mixture was then filtered and the filtrate was concentrated to yielda residue which was purified by column chromatography (EA/PE=1/1 to pureEA) to give the compound as a colorless oil (500 mg, 75% purity). Andused directly in the next step. LCMS (m/z): 190 (M+1).

Step 2: tert-Butylcyclopentyl(3-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)benzyl)carbamate

To a solution of 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline(170 mg, 0.899 mmol) in EtOH (10 mL) was added tert-butyl3-aminobenzyl(cyclopentyl)carbamate (313 mg, 1.08 mmol) and the solutionheated at 100° C. in a sealed tube overnight. The solvent was thenevaporated off and the residue purified by prep-TLC to give 100 mgcompound as colorless oil (280 mg, 65%). LCMS (m/z): 480.3 (M+1)

Step 3:1-((3-((cyclopentylamino)methyl)phenyl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of tert-butylcyclopentyl(3-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)benzyl)carbamate(100 mg, 0.208 mmol) in EA (15 mL) was added TFA (5 mL). The reactionsolution was stirred at reflux for 1 h. Once cooled, the solvent wasremoved by concentration and the crude product was purified bypreparative HPLC separation to get the final compound as the TFA salt(32 mg, 45% yield).

¹H NMR (400 MHz, MeOD): δ 8.47 (s, 2H), 7.28-7.10 (m, 5H), 6.86 (s, 1H),6.83-6.73 (m, 2H), 4.29 (br. s, 1H), 4.25 (s, 2H), 3.58 (br. s, 1H),3.41-3.31 (m, 2H), 3.30-3.04 (m, 6H), 2.16 (br. s, 2H), 1.84 (br. s,2H), 1.69 (br. s, 4H). LCMS (m/z): 380.3 (M+1)

Compound 43 1-([1,1′-biphenyl]-3-yloxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl) propan-2-ol

Step 1: 2-((3-bromophenoxy)methyl) oxirane

To a solution of NaH (416.2 g, 17.34 mmol) in DMF (10 mL) was added3-bromophenol (1.0 g, 5.78 mmol) at 9° C. and stirred for 5 minutes. Tothis mixture was added 2-bromophenol (1.2 g, 8.67 mmol) at 9° C. Thereaction mixture was stirred for a further 16 h at 9° C. TLC (PE:EA=5:1)showed that the reaction was completed. The mixture was treated withwater (50 mL) and extracted with EA (2×20 mL). The organic layer waswashed with NaHCO₃, brine (30 mL) then dried over Na₂SO₄ andconcentrated to give 2-((3-bromophenoxy)methyl) oxirane (1.5 g, crude)as colorless oil which was used in next step without furtherpurification. LCMS (m/z): 229.1/230.1 [M+H]⁺/[M+2H]⁺

Step 2: 1-(3-bromophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of 2-((3-bromophenoxy) methyl) oxirane (1.5 g, 6.55 mmol)in MeOH (15 mL) were added 1,2,3,4-tetrahydroisoquinoline (1.0 g, 7.86mmol) at 8° C. The mixture was refluxed for 16 h. TLC (PE:EA=2:1) showedthat the reaction was completed and the mixture was concentrated toyield crude material which was purified using column chromatography onsilica gel to give the desired compound (1.8 g, 78.3%) as colorless oil.LCMS (m/z): 362.1/363.1 [M+H]⁺/[M+2H]⁺

Step 3: 1-([1, 1′-biphenyl]-3-yloxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl) propan-2-ol

To a solution of1-(3-bromophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (300mg, 0.828 mmol) in dioxane (4 ml) and H₂O (1 mL) was added phenylboronicacid (151.5 mg, 1.24 mmol), Pd(dppf)Cl₂ (30.3 g, 0.041 mmol) and K₂CO₃(343.4 g, 2.484 mmol) at 6° C. The reaction mixture was stirred for 16 hat 80° C. It was concentrated to remove the solvents and the residue wasdissolved in ethyl acetate, washed with water. The separated organiclayer was concentrated and the crude product purified by HPLC separationto give the title compound (89 mg, 30.0%) as a white solid. ¹HNMR(CH₃OD, 400 MHz) δ: 8.47 (br, 1H), 7.60-7.62 (m, 2H), 7.40-7.46 (m, 2H),7.33-7.38 (m, 2H), 7.17-7.28 (m, 6H), 6.97-6.99 (m, 1H), 4.46-4.50 (m,1H), 4.35 (s, 2H), 4.12 (d, J=2.6, 2H), 3.47-3.50 (m, 2H), 3.26-3.37 (m,2H), 3.16 (s, 2H). LCMS (m/z): 360.2 [M+H]⁺

Compound 451-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

To a solution of1-(3-bromophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (200mg, 0.55 mmol) in a mixed solution (Dioxane/H₂O=4/1 mL) were added1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole(214 mg, 0.83 mmol), Pd(dppf)Cl₂ (40 mg, 0.06 mmol) and Cs₂CO₃ (360 mg,1.10 mmol). The reaction mixture was heated at 120° C. under microwavecondition for 40 min. The solvent was removed by concentration and theresidue was dissolved in ethyl acetate, washed with water. The separatedorganic layer was concentrated and the crude product was purified bypreparative HPLC separation to give the title compound (60 mg, yield26%) MS (ESI⁺) e/z: 414.1 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 9.45(s, 1H), 8.21 (s, 1H), 8.00-7.93 (m, 2H), 7.50-7.24 (m, 7H), 7.10-7.07(m, 1H), 4.76-4.70 (m, 1H), 4.62-4.49 (m, 2H), 4.24 (s, 3H), 4.22-4.19(m, 2H), 3.98-3.94 (m, 1H), 3.60-3.51 (m, 3H), 3.41-3.21 (m, 2H).

Compound 581-(3-(1H-indazol-6-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of 6-bromo-1H-indazole (64.2 g, 0.326 mmol) in dioxane (4ml) and water (1 mL) was added1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol (200 mg, 0.489 mmol), Pd(dppf)Cl₂ (24 mg, 0.03257mmol) and K₂CO₃ (135 mg, 0.98 mmol) at 16° C. The reaction mixture wasstirred for 16 h at 100° C. TLC (PE:EA=1:1) showed that the reaction wascompleted. The mixture was concentrated to provide a crude product whichwas purified by HPLC separation to give the title compound (22.0 g,11.3%) as a white solid. ¹HNMR (CD₃OD, 400 MHz) δ: 8.49 (br, 1H), 8.07(s, 1H), 7.85-7.83 (m, 1H), 7.76-7.71 (m, 1H), 7.44-7.10 (m, 8H),7.03-6.98 (m, 1H), 4.50-4.42 (m, 1H), 4.30 (s, 2H), 4.15-4.13 (m, 2H),3.49-3.40 (m, 2H), 3.36-3.31 (m, 2H), 3.19-3.10 (m, 2H). LCMS (m/z):400.2 [M+H]⁺

Compound 591-(3-(benzo[d][1,3]dioxol-5-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of 5-bromobenzo[d][1,3]dioxole (100 mg, 0.49 mmol) indioxane (4 ml) and H₂O (1 mL) was added1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(300 mg, 0.73 mmol), Pd(dppf)Cl₂ (36 mg, 0.049 mmol) and K₂CO₃ (203.2 g,1.47 mmol) at 12° C. The reaction mixture was stirred for 16 h at 100°C. Upon completion, the reaction mixture was concentrated to get thecrude material which was purified by HPLC separation to give the titlecompound (146 mg, 73.7%) as a white solid. ¹HNMR (CH₃OD, 400 MHz) δ:8.39 (br, 1H), 7.48-7.10 (m, 9H), 6.95-6.86 (m, 2H), 6.00 (s, 2H),4.56-4.39 (m, 3H), 4.16-4.08 (m, 2H), 3.60-3.51 (m, 2H), 3.48-3.31 (m,2H), 3.23-3.11 (m, 2H). LCMS (m/z): 404.2 [M+H]⁺

Compound 601-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-methyl-1H-benzo[d]imidazol-5-yl)phenoxy)propan-2-ol

To a solution of 5-bromo-1-methyl-1H-benzo[d]imidazole (69 mg, 0.3257mmol) in dioxane (4 ml) and H₂O (1 mL) was added1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(200 mg, 0.4886 mmol), Pd(dppf)Cl₂ (24 mg, 0.03257 mmol) and K₂CO₃ (135mg, 0.98 mmol) at 16° C. The reaction mixture was stirred for 16 h at100° C. and the reaction was shown to be complete by TLC. The mixturewas concentrated to get the crude which was purified by HPLC separationto give the formate salt of title compound (30.0 g, 14.9%) as a whitesolid. ¹HNMR (CH₃OD, 400 MHz) δ: 8.48 (br, 1H), 8.20-8.19 (m, 1H),7.90-7.72 (m, 1H), 7.63 (s, 1H), 7.45-7.12 (m, 7H), 7.13-6.95 (m, 1H),4.49-4.40 (m, 1H), 4.25-4.11 (m, 2H), 3.95 (s, 2H), 3.38-3.29 (m, 2H),3.28-3.05 (m, 4H). LCMS (m/z): 414.2 [M+H]⁺

Compound 621-((2′-chloro-5′-(trifluoromethyl)-[1,1′-biphenyl]-3-yl)oxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

A mixture of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(200 mg, 0.489 mmol), 2-bromo-1-chloro-4-(trifluoromethyl)benzene (126mg, 0,489 mmol), Pd(dppf)Cl₂ (36 mg, 0.049 mmol), K₂CO₃ (202 mg, 1.47mmol) in H₂O-dioxane (1 mL/3 mL) was stirred at 100° C. under microwaveheating for 15 min. The solvent was removed and the crude productpurified by HPLC separation to give the title compound as the TFA salt(186 mg, 85%)¹HNMR (CH₃OD, 400 MHz) δ: 7.76-7.63 (m, 3H), 7.46-7.40 (m,1H), 7.38-7.20 (m, 4H), 7.11-7.02 (m, 3H), 4.78-4.40 (m, 3H), 4.18-4.08(m, 2H), 4.02-3.84 (m, 1H), 3.67-3.40 (m, 3H), 3.38-3.16 (m, 2H). LCMS(m/z): 462.2 [M+H]⁺

Compound 651-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(quinolin-8-yl)phenoxy)propan-2-ol

A mixture of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(200 mg, 0.489 mmol), 8-bromoquinoline (102 mg, 0.489 mmol), Pd(dppf)Cl₂(36 mg, 0.049 mmol), K₂CO₃ (202 mg, 1.47 mmol) in H₂O-dioxane (1 mL/3mL) was stirred at 100° C. under microwave heating conditions for 15min. The solvent was removed by concentration and the crude productpurified by HPLC separation to give the title compound as the TFA salt(93 mg, 46.3%). ¹HNMR (CH₃OD, 400 MHz) δ: 9.14-9.10 (m, 1H), 9.05-9.01(m, 1H), 8.36-8.30 (m, 1H), 8.08-7.96 (m, 3H), 7.60-7.52 (m, 1H),7.36-7.18 (m, 7H), 4.77-4.40 (m, 3H), 4.20-4.12 (m, 2H), 4.00-3.80 (m,1H), 3.60-3.48 (m, 3H), 3.38-3.16 (m, 2H). LCMS (m/z): 411.2 [M+H]⁺

Compound 681-((2′,3′-difluoro-[1,1′-biphenyl]-3-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

A mixture of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)amino)propan-2-ol(200 mg, 0.489 mmol), 1-chloro-2,3-difluorobenzene (94 mg, 0,489 mmol),Pd(dppf)Cl₂ (36 mg, 0.049 mmol), K₂CO₃ (202 mg, 1.47 mmol) inH₂O-dioxane (1 mL/3 mL) was stirred at 100° C. under microwave heatingfor 15 min. The solvent was removed and concentrated to yield a crudeproduct which was purified by HPLC separation to give the title compoundas the formate salt (66 mg, 34%). ¹H NMR (400 MHz, MeOD): δ 8.39 (s,1H), 7.26 (t, J=8.0 Hz, 1H), 7.32-7.16 (m, 9H), 7.08-7.05 (m, 1H),4.55-4.49 (m, 1H), 4.46 (s, 2H), 4.14-4.12 (m, 2H), 3.60 (t, J=6.4 Hz,2H), 3.34-3.33 (m, 2H), 3.23-2.19 (m, 2H), ppm; ESI-MS (m/z): 396.2[M+1]⁺.

Compound 721-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((3′-(hydroxymethyl)-[1,1′-biphenyl]-3-yl)oxy)propan-2-ol

To a solution of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(220 mg, 0.538 mmol) in dioxane/H₂O (1:1) (3 mL) was added(3-bromophenyl)methanol (100 mg, 0.538 mmol), Pd(dppf)Cl₂ (20 mg, 0.0269mmol) and K₃PO₄ (342 mg, 1.613 mmol). The reaction mixture was stirredat 100° C. under N₂ for 12 h. After cooling, the reaction mixture wasextracted with DCM, H₂O and the separated organic layer dried oversodium sulfate before being filtered and concentrated. The resultingcrude product was purification by prep-HPLC to afford the desiredproduct as the formate salt (80 mg, 39% yield). ¹H NMR (400 MHz, MeOD):δ 8.49 (br. s, 1H), 7.62 (s, 1H), 7.51 (d, J=7.6 Hz, 1H) 7.44-7.18 (m,9H), 6.99 (dd, J=7.6 Hz, J₂=2 Hz, 1H), 4.69 (s, 2H), 4.52-4.46 (m, 1H),4.39 (s, 2H), 4.16 (d, J=4.8 Hz, 2H), 3.54 (t, J=6.4 Hz, 2H), 3.10-3.13(m, 4H). LCMS (m/z): 390.2 (M+1).

Compound 751-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4′-methoxy-[1,1′-biphenyl]-3-yl)oxy)propan-2-ol

To a mixture of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(250 mg, 0.61 mmol) in Dioxane (3 mL) was added 4-bromoanisole (115 mg,0.61 mmol), 2N K₂CO₃ (1 mL, 2 mmol) and PdCl₂(dppf)₂ (20 mg, 0.025mmol). The reaction mixture was heated at 120° C. under microwaveheating conditions for 30 minutes before concentrated to remove thesolvent. The residue was dissolved in ethyl acetate, washed with waterwith the separated organic layer dried and concentrated to yield a crudeproduct which was purified by prep-HPLC separation to give titlecompound (116 mg, 49%).

¹H NMR (400 MHz, MeOD): δ 7.55-7.51 (m, 2H), 7.35-7.27 (m, 4H),7.24-7.15 (m, 3H), 7.01-6.97 (m, 2H), 6.92-6.89 (m, 1H), 4.66 (brs, 1H),4.56-4.51 (m, 2H), 4.16-4.07 (m, 2H), 3.92 (brs, 1H), 3.83 (s, 3H),3.58-3.48 (m, 3H), 3.32-3.22 (m, 2H), ppm; ESI-MS (m/z): 390.2 [M+1]⁺.

Compound 821-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-(4-methoxybenzyl)-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

Step 1: 5-bromo-N-(4-methoxybenzyl)-2-nitroaniline

To a solution of 4-bromo-2-fluoro-1-nitrobenzene (500 mg, 2.27 mmol) inDMF (6 mL) was added (4-methoxyphenyl)methanamine (342 mg, 2.5 mmol) andTEA (345 mg, 3.4 mmol). The reaction mixture was then stirred at 120° C.for 30 min under microwave heating. After cooling, the mixture wasdiluted with water (40 mL) and the resulting precipitate collected byfiltration, washed with water and dried in vacuo to give the desiredproduct (750 mg, crude, 97% yield) which was used in next step withoutfurther purification. LCMS (m/z): 338.1[M+H]⁺

Step 2: 5-bromo-N-(4-methoxybenzyl)benzene-1,2-diamine

To a solution of 5-bromo-N-(4-methoxybenzyl)-2-nitroaniline (750 mg,crude) in EtOH (8 mL) and H₂O (8 mL) was added Fe powder (766 mg, 13.7mmol) and ammonium chloride (733 mg, 13.7 mmol). The mixture was stirredat 60° C. for 4 h then filtered and the filtrate was concentrated toremove EtOH. The residue was then diluted with water and extracted withEtOAc. The organic layer was concentrated to give the desired product(550 mg, crude, 82% yield) with the crude product being used in nextstep without further purification. LCMS (m/z): 308.1 [M+H]⁺

Step 3: 6-bromo-1-(4-methoxybenzyl)-1H-benzo[d]imidazole

To a solution of 5-bromo-N-(4-methoxybenzyl)benzene-1,2-diamine (550 mg,crude) in HC(OMe)₃ (20 mL) was added TsOH.H₂O (30 mg, 0.16 mmol). Themixture was stirred at 100° C. for 4 h. The reaction solution was thenconcentrated and the residue was washed with water and extracted withEtOAc. The organic layer was concentrated to give the desired product(450 mg, 79% yield) and the crude product used in next step withoutfurther purification. LCMS (m/z): 318.1 [M+H]⁺

Step 4:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-(4-methoxybenzyl)-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

A mixture of 6-bromo-1-(4-methoxybenzyl)-1H-benzo[d]imidazole (200 mg,0.63 mmol), 1-(3, 4-dihydroisoquinolin-2-(1H)-yl)-3-(3-(4, 4, 5,5-tetramethyl-1, 3, 2-dioxaborolan-2-yl)phenoxy)propan-2-ol (284 mg,0.69 mmol), K₂CO₃ (261 mg, 1.89 mmol) and Pd(dppf)Cl₂ (50 mg) in dioxane(8 mL) and H₂O (2 mL) was stirred at 100° C. for 16 h. The catalyst wasfiltered and the filtrate concentrated. The residue was then purified byPrep-HPLC to give the title compound (79 mg, 24.2%). ¹HNMR (CH₃OD, 400MHz) δ: 8.39 (br, 1H), 8.25 (s, 1H), 7.73-7.68 (m, 1H), 7.64-7.61 (m,1H), 7.56-7.48 (m, 1H), 7.48-7.42 (m, 1H), 7.28-7.14 (m, 8H), 6.97-6.85(m, 3H), 5.44 (s, 2H), 4.50-4.45 (m, 1H), 4.35 (s, 2H), 4.10-4.09 (m,2H), 3.70 (s, 3H), 3.51-3.48 (m, 2H), 3.36-3.31 (m, 2H), 3.19-3.10 (m,2H). LCMS (m/z): 520.2 [M+H]⁺

Compound 841-(3-(1H-benzo[d][1,2,3]triazol-6-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

Step 1: 6-bromo-1H-benzo[d][1,2,3]triazole

The mixture of 4-bromobenzene-1,2-diamine (200 mg, 1.1 mmol) and NaNO₂(569 mg, 5.3 mmol) in H₂O/AcOH (5 mL/50 mL) was reacted at 80° C. for 6hours. The mixture was concentrated, the residue was dissolved in DCMand extracted with water, and the organic phase dried by Na₂SO₄ andconcentrated to give 300 mg of crude product which was used withoutfurther purification. LCMS (m/z): 198.1/199.1 [M+H]⁺/[M+2H]⁺

Step 2:1-(3-(1H-benzo[d][1,2,3]triazol-6-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a mixture of 6-bromo-1H-benzo[d][1,2,3]triazole (300 mg, crude) inTHF (50 mL) was added NaH (50 mg, 1.25 mmol) at 0° C., and the mixturewas stirred at r.t, for 30 min. The SEM-Cl (100 mg, 1.1 mmol) was addedto the mixture and the mixture was stirred at r.t. for 1 hour. Themixture was quenched by water and then extracted with EA, the organicphases was concentrated. The residue was mixed with Pd(dppf)Cl₂ (36 mg,0.049 mmol), K₂CO₃ (202 mg, 1.47 mmol) in H₂O-dioxane (1 mL/3 mL) wasstirred at 100° C. over microwave for 15 min. The solvent was removed byconcentration and the residue was dissolved in DCM, HCl/EA (4 M) wasadded to the mixture, then the mixture was stirred at r.t. for 1 hour.The mixture was concentrated and the crude product was purified by HPLCseparation (22 mg, 5% overall). ¹HNMR (CH₃OD, 400 MHz) δ: 8.83 (br, 1H),8.05 (s, 1H), 7.96-7.93 (m, 1H), 7.78-7.73 (m, 1H), 7.45-7.18 (m, 7H),7.04-7.02 (m, 1H), 4.53-4.48 (m, 1H), 4.38 (s, 2H), 4.18-4.16 (m, 2H),3.54-3.51 (m, 2H), 3.39-3.38 (m, 2H), 3.21-3.13 (m, 2H). LCMS (m/z):401.2 [M+H]⁺

Compound 851-(3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of 6-bromo-2,3-dihydrobenzo[b][1,4]dioxine (150 mg, 0.7mmol) in dioxane (4 ml) and H₂O (1 mL) was added1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(428.3 g, 1.05 mmol), Pd(dppf)Cl₂ (51.2 g, 0.07 mmol) and K₂CO₃ (290.3g, 2.1 mmol) at 16° C. The reaction mixture was stirred for 16 h at 100°C. until TLC indicated the reaction was completed. The mixture wasconcentrated to get yield a crude which was purified by HPLC separationto give the formate salt of the title compound (136.0 g, 46.5%) as awhite solid. ¹HNMR (CH₃OD, 400 MHz) δ: 8.40 (br, 1H), 7.35-7.06 (m, 9H),6.95-6.49 (m, 2H), 4.51-4.98 (m, 1H), 4.40 (s, 2H), 4.28 (s, 4H),4.14-4.08 (m, 2H), 3.57-3.51 (m, 2H), 3.40-3.29 (m, 2H), 3.21-3.11 (m,2H). LCMS (m/z): 418.2 [M+H]⁺

Compound 861-(3-(dibenzo[b,d]furan-4-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of1-(3-bromophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (300mg, 0.828 mmol) in dioxane (4 ml) and H₂O (1 mL) was addeddibenzo[b,d]furan-4-ylboronic acid (212 mg, 1.0 mmol), Pd(dppf)Cl₂ (60.6g, 0.083 mmol) and K₂CO₃ (343.3 g, 2.48 mmol) at 12° C. The reactionmixture was stirred for 16 h at 100° C. TLC (PE:EA=1:1) showed that thereaction was completed. The mixture was concentrated to get the crudewhich was purified by HPLC separation to give the formate salt of thetitle compound (181.0 g, 48.7%) as a white solid. ¹HNMR (CH₃OD, 400 MHz)δ: 8.42 (br, 1H), 8.11-8.00 (m, 2H), 7.68-7.34 (m, 8H), 7.29-7.16 (m,4H), 7.08-7.02 (m, 1H), 4.56-4.48 (m, 1H), 4.46-4.32 (m, 2H), 4.20-4.12(m, 2H), 3.58-3.48 (m, 2H), 3.42-3.36 (m, 2H), 3.21-3.09 (m, 2H). LCMS(m/z): 450.2 [M+H]⁺

Compound 891-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(quinolin-3-yl)phenoxy)propan-2-ol

To a solution of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(250 mg, 0.61 mmol) in Dioxane (3 mL) was added 3-bromoquinoline (127mg, 0.61 mmol), 2N K₂CO₃ (1 mL, 2 mmol) and PdCl₂(dppf)₂ (20 mg, 0.025mmol). The reaction mixture was heated at 120° C. under microwaveconditions for 30 min. the mixture was then concentrated to remove thesolvents, the residue dissolved in ethyl acetate and washed with water.The separated organic layer dried and concentrated to yield the crudeproduct which was purified HPLC separation to give the title compound(133 mg, 53.2%).

¹HNMR (CH₃OD, 400 MHz) δ: 9.39 (s, 1H), 9.12-9.08 (m, 1H), 8.28-8.19 (m,2H), 8.06-8.00 (m, 1H), 7.90-7.84 (m, 1H), 7.88-7.46 (m, 3H), 7.36-7.20(m, 4H), 7.18-7.12 (m, 1H), 4.78-4.40 (m, 3H), 4.23-4.16 (m, 2H),4.02-4.80 (m, 1H), 3.65-3.40 (m, 3H), 3.40-3.20 (m, 2H). LCMS (m/z):411.2 [M+H]⁺

Compound 1021-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)amino)propan-2-ol

Step 1:1-((3-bromophenyl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a stirred mixture of 3-bromoaniline (300 mg, 1.75 mmol) in EtOH (10mL) was added 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (331mg, 1.75 mmol). The mixture was stirred at 80° C. for 16 hours. Thereaction mixture was concentrated. The residue was purified by prep-TLCto afford the desired compound (120 mg). LCMS (m/z): 361.1/362.1[M+H]⁺/[M+2H]⁺

Step 2:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)amino)propan-2-ol

To a stirred mixture of1-((3-bromophenyl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl) propan-2-ol(120 mg, 0.33 mmol) in dioxane:H₂O (15 mL, 2:1) was added1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole(93.6 g, 0.33 mmol), Cs₂CO₃ (323 mg, 0.99 mmol) and then Pd(dppf)Cl₂ (10mg). The mixture was degassed by N₂ for 4 times and then stirred at 100°C. for 16 hours. The reaction mixture was quenched with water (30 mL),extracted with EA (30 mL×3). The combined extracts were washed withbrine (30 mL), dried over anhydrous Na₂SO₄ and concentrated. The residuewas purified by prep-HPLC to afford the title compound (46 mg, 34%).¹HNMR (CDCl₃, 400 MHz) δ: 7.87 (s, 1H), 7.83-7.81 (m, 1H), 7.55-7.52 (m,2H), 7.29-7.25 (m, 1H), 7.15-7.10 (m, 3H), 7.04-6.98 (m, 2H), 6.92 (s,1H), 6.66-6.64 (m, 1H), 4.18-4.11 (m, 1H), 3.89-3.83 (m, 4H), 3.70-3.64(m, 1H), 3.42-3.38 (m, 1H), 3.21-3.14 (m, 1H), 3.02-2.94 (m, 3H),2.71-2.61 (m, 3H). LCMS (m/z): 413.2 [M+H]⁺

Compound 1031-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

Step 1: N-(5-bromo-2-nitrophenyl)tetrahydro-2H-pyran-4-amine

To a solution of 4-bromo-2-fluoro-1-nitrobenzene (2 g, 9.137 mmol) inDMF (10 mL) was added tetrahydro-2H-pyran-4-amine (1.108 g, 10.964 mmol)and TEA (2.772 g, 27.410 mmol). The reaction mixture was heated at 120°C. in microwave reactor for 30 mins. The reaction mixture was dilutedwith H₂O then extracted with EA. The combined organic layers were driedover sodium sulfate, filtered and concentrated to yield a crude product(2.5 g) which was used in next step without further purification. LCMS(m/z): 301.0 (M+1).

Step 2: 5-Bromo-N1-(tetrahydro-2H-pyran-4-yl)benzene-1,2-diamine

To a solution of N-(5-bromo-2-nitrophenyl)tetrahydro-2H-pyran-4-amine(2.5 g, 8.333 mmol) in EtOH/H₂O (1:1) (50 mL) was added iron powder (2.8g, 50 mmol) and NH₄Cl (2.65 g, 50 mmol). The reaction mixture was heatedat 60° C. for 12 h. After filtering the reaction mixture, the resultingfiltrate was extracted with DCM with the separated organic layer driedover sodium sulfate, filtered and concentrated to obtain crude productwhich was used in next step without further purification.

Step 3: 6-Bromo-1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazole

To a solution of compound5-bromo-N1-(tetrahydro-2H-pyran-4-yl)benzene-1,2-diamine (500 mg, 1.852mmol) in HC(OMe)₃ (2 mL) was added TsOH (15 mg, 0.0789 mmol). Thereaction mixture was stirred at 100° C. for 12 h. The reaction mixturewas extracted with DCM washed with water and the organic layer driedover sodium sulfate which was removed by filtration. After concentrationof the filtrate, the crude product was purified by column (201 mg,38.53%). LCMS (m/z): 281.0, 283.0 (M+1).

Step 4: 2-((3-bromophenoxy)methyl)oxirane

To a solution of compound NaH (416.2 g, 17.34 mmol) in DMF (10 mL) wasadded 3-bromophenol (1.0 g, 5.78 mmol) at 20° C. After addition thesolution was stirred for 5 minutes at this temperature before theaddition of 2-(bromomethyl)oxirane (1.2 g, 8.67 mmol). The reactionmixture was stirred for 16 h at 20° C. Once complete the reactionmixture was treated with water (50 mL) and extracted with EA (2×20 mL)and the organic layers combined and washed with NaHCO₃, brine (30 mL),dried over Na₂SO₄ and concentrated to give the compound2-((3-bromophenoxy)methyl)oxirane (1.5 g, crude) as colorless oil whichwas used in next step without further purification.

Step 5:1-(3-bromophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of 2-((3-bromophenoxy)methyl)oxirane (1.5 g, 6.55 mmol) inMeOH (15 mL) was added 1,2,3,4-tetrahydroisoquinoline (1.0 g, 7.86 mmol)at 20° C. The mixture was heated at reflux temperature for 16 h, untilthe reaction was shown to be complete. The mixture then was concentratedto provide the crude which was purified using column chromatography onsilica gel producing the desired1-(3-bromophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (1.8g, 78.3%) as colorless oil. LCMS (m/z): 362.1 (M+1).

Step 6:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol

To a solution of1-(3-bromophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (1.31g, 4.01 mmol) in DMSO (10 ml) was added4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (1.2 g, 4.42mmol), KOAc (1.2 g, 12.04 mmol) and Pd(dppfCl)₂ (0.88 g, 1.2 mmol) at20° C. The mixture was stirred for 4 hours at 100° C. until the reactionwas complete. The mixture was then treated with water (50 ml) andextracted with EA (2×20 ml) and the combined organic layers dried andconcentrated to get the crude which was purified by flash column to givethe desired1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(795 mg, 48.5%) as a colorless oil. LCMS (m/z): 410.2 (M+1).

Step 7:1-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-(3-(1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

To a solution of6-bromo-1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazole (200 mg, 0.714mmol) in dioxane/H₂O (1:1) (8 mL) were added1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(292 mg, 0.714 mmol) Pd(dppf)Cl₂ (26 mg, 0.0357 mmol) and K₃PO₄ (454 mg,2.143 mmol). The reaction mixture was stirred at 100° C. under N₂ for 12h. After cooling, the reaction mixture was extracted with DCM and water.The combined organic layers were dried over sodium sulfate, filtered andconcentrated to yield a crude product which was purified by prep-HPLC toafford the title compound the TFA salt (38 mg, Yield 11.2%). ¹H NMR (400MHz, MeOD): δ 8.39 (s, 1H), 8.36 (s, 1H), 7.87 (s, 1H), 7.75 (d, J=8.4Hz, 1H), 7.57 (d, J=8.4 Hz, 1H), 7.42-7.18 (m, 7H), 6.99 (d, J=8.0 Hz,1H), 4.79-4.71 (m, 1H), 4.55-4.52 (m, 1H), 4.46 (s, 2H), 4.19-4.12 (m,4H), 3.70 (t, J=11.2 Hz, 2H), 3.61 (t, J=6.4 Hz, 2H), 3.47-3.37 (m, 2H),3.33 (s, 2H), 3.22-3.18 (m, 2H), 2.28-2.13 (m, 4H). LCMS (m/z): 484.2(M+1).

Compound 1041-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-phenyl-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

Step 1: 5-bromo-2-nitro-N-phenylaniline

Aniline (930 mg, 10.0 mmol) was added to a solution of DIPEA (1.94 g,15.0 mmol) and 4-bromo-2-fluoro-1-nitrobenzene (2.2 g, 10.0 mmol) inDMSO (16 mL). The reaction mixture was stirred at 60° C. for 4 h. Uponcompletion, the mixture was diluted with water (50 mL), and theresulting precipitate collected by filtration, washed with water anddried in vacuo. The crude product was used in next step without furtherpurification. LCMS (m/z): 293.1/294.1 [M+H]⁺/[M+2H]⁺

Step 2: 5-bromo-N1-phenylbenzene-1,2-diamine

To a solution of 5-bromo-2-nitro-N-phenylaniline (1.0 g, 3.41 mmol) inEtOH (15 mL) and H₂O (15 mL) was added iron powder (1.15 g, 20.5 mmol)and NH₄Cl (1.09 g, 20.5 mmol). The mixture was stirred at 60° C. for 4h. After completion, the reaction mixture was filtered and the filtrateconcentrated to remove EtOH. The residue was diluted with water andextracted with EtOAc. The organic layer was concentrated to give thecrude diamine product which was used in next step without furtherpurification. LCMS (m/z): 263.1/264.1 [M+H]⁺/[M+2H]⁺

Step 3: 6-Bromo-1-phenyl-1H-benzo[d]imidazole

To a solution of 5-bromo-N1-phenylbenzene-1,2-diamine (650 mg, 2.47mmol) in HC(OMe)₃ (15 mL) was added TsOH—H₂O (38 mg, 0.2 mmol). Themixture was stirred at 100° C. for 4 h. Upon completion, the reactionsolution was concentrated, the residue was extracted with EtOAc andwashed with water. The organic layer was concentrated to give the crude6-bromo-1-phenyl-1H-benzo[d] imidazole which was used in next stepwithout further purification. LCMS (m/z): 273.1/274.1 [M+H]⁺/[M+2H]⁺

Step 4:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-phenyl-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

A mixture of 6-bromo-1-phenyl-1H-benzo[d] imidazole (200 mg, 0.73 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(330 mg, 0.81 mmol), K₂CO₃ (303 mg, 2.19 mmol) and Pd(dppf)Cl₂ (50 mg)in a solution of dioxane (8 mL) and H₂O (2 mL) was stirred at 100° C.for 16 h. The catalyst was filtered, and the filtrate concentrated. Theresidue was purified by prep-HPLC to give the desired title compound(152 mg, 43.8%). ¹HNMR (CH₃OD, 400 MHz) δ: 8.42 (s, 1H), 8.37 (br, 1H),8.22-8.20 (m, 1H), 7.70-7.52 (m, 7H), 7.49-7.18 (m, 7H), 6.96-6.93 (m,1H), 4.53-4.38 (m, 3H), 4.02-3.96 (m, 2H), 3.61-3.55 (m, 2H), 3.48-3.36(m, 2H), 3.25-3.13 (m, 2H). LCMS (m/z): 476.2 [M+H]⁺

Compound 1051-(3-(1-benzyl-1H-benzo[d]imidazol-6-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

Step 1: N-benzyl-5-bromo-2-nitroaniline

To a solution of 4-bromo-2-fluoro-1-nitrobenzene (500 mg, 2.27 mmol) inDMF (6 mL) were added phenylmethanamine (268 mg, 2.5 mmol) and TEA (345mg, 3.4 mmol). The reaction mixture was stirred at 120° C. for 30 minunder microwave. The reaction solution was diluted with water (40 mL),and the resulting precipitate was collected by filtration, washed withwater and dried in vacuo. The crude product was used in next stepwithout further purification. LCMS (m/z): 307.1/308.1 [M+H]⁺/[M+2H]⁺

Step 2: N1-benzyl-5-bromobenzene-1,2-diamine

To a solution of N-benzyl-5-bromo-2-nitroaniline (700 mg, 2.28 mmol) inEtOH (8 mL) and water (8 mL) was added iron powder (766 mg, 13.7 mmol)and NH₄Cl (733 mg, 13.7 mmol). The mixture was stirred at 60° C. for 4h. The reaction solution was filtered with the filtrate concentrated toremove EtOH. The residue was diluted with water and extracted with EtOAcwith the organic layer concentrated to give the crudeN1-benzyl-5-bromobenzene-1,2-diamine. The crude product was used in nextstep without further purification. LCMS (m/z): 277.1/278.1[M+H]⁺/[M+2H]⁺

Step 3: 1-benzyl-6-bromo-1H-benzo[d]imidazole

To a solution of N1-benzyl-5-bromobenzene-1,2-diamine (450 mg, 1.63mmol) in HC(OMe)₃ (15 mL) was added TsOH—H₂O (30 mg, 0.16 mmol). Themixture was stirred at 100° C. for 4 h. The reaction solution wasconcentrated, and the residue was washed with water and extracted withEtOAc. The organic layer was concentrated to give the crude1-benzyl-6-bromo-1H-benzo [d]imidazole. The crude product was used innext step without further purification. LCMS (m/z): 287.1/288.1[M+H]⁺/[M+2H]⁺

Step 4:1-(3-(1-benzyl-1H-benzo[d]imidazol-6-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

A mixture of 1-benzyl-6-bromo-1H-benzo[d]imidazole (200 mg, 0.70 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(314 mg, 0.77 mmol), K₂CO₃ (290 mg, 2.1 mmol) and Pd(dppf)Cl₂ (50 mg) indioxane (8 mL) and H₂O (2 mL) was stirred at 100° C. for 16 h. Thecatalyst was filtered and the filtrate was concentrated. The residue waspurified by prep-HPLC to give the title compound (108 mg, 31.6%). ¹HNMR(CH₃OD, 400 MHz) δ: 8.43 (br, 1H), 8.25 (s, 1H), 7.75-7.68 (m, 1H),7.59-7.48 (m, 2H), 7.33-7.10 (m, 12H), 6.95-6.89 (m, 1H), 5.46 (s, 2H),4.52-4.46 (m, 1H), 4.43-4.32 (m, 2H), 4.12-4.01 (m, 2H), 3.57-3.47 (m,2H), 3.40-3.25 (m, 2H), 3.19-3.04 (m, 2H). LCMS (m/z): 490.2 [M+H]⁺

Compound 1095-(3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)-1H-benzo[d]imidazol-2(3H)-one

Step 1: 5-Bromo-1H-benzo[d]imidazol-2(3H)-one

The mixture of 4-bromobenzene-1,2-diamine (200 mg, 1.1 mmol), CDI (174mg, 1.1 mmol) in dioxane was reacted at 50° C. for 16 hours. The mixturewas concentrated and the residue was dissolved in DCM and extracted withwater with the combined organic phases dried by Na₂SO₄ and concentratedto give 300 mg of crude product which was used without furtherpurification. LCMS (m/z): 213.1/214.1 [M+H]⁺/[M+2H]⁺

Step 2:5-(3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)-1H-benzo[d]imidazol-2(3H)-one

A mixture of 5-bromo-1H-benzo[d]imidazol-2(3H)-one (200 mg, 0.489 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(100 mg), Pd(dppf)Cl₂ (36 mg, 0.049 mmol), K₂CO₃ (202 mg, 1.47 mmol) inH₂O-dioxane (1 mL/3 mL) was stirred and heated in am microwave reactorto 100° C. for 15 min. Once cooled, the solvent was removed byconcentration and the crude product purified by HPLC separation to givethe title compound (74 mg, 36.5%). ¹HNMR (CH₃OD, 400 MHz) δ: 7.49-7.10(m, 10H), 6.98-6.92 (m, 1H), 4.76-4.42 (m, 3H), 4.20-4.09 (m, 2H),3.98-3.89 (m, 1H), 3.62-3.48 (m, 3H), 3.36-3.14 (m, 2H). LCMS (m/z):416.2 [M+H]⁺

Compound 1131-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4-methylpyridin-3-yl)phenoxy)propan-2-ol

To a mixture of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)amino)propan-2-ol(84.05 g, 0.4886 mmol) in dioxane (4 ml) and H₂O (1 mL) was addedcompound 3-bromo-4-methylpyridine (300 mg, 0.733 mmol), Pd(dppf)Cl₂ (36mg, 0.049 mmol) and K₂CO₃ (202.6 g, 1.466 mmol) at 14° C. The reactionmixture was stirred for 16 h at 100° C. until the reaction looked to becomplete by TLC showed. The mixture was then concentrated to get the acrude material which was purified by HPLC separation to give the titlecompound as the formate salt (78.0 g, 28.4%) as a white solid. ¹H NMR(400 MHz, MeOD): 8.40 (s, 2H), 8.34 (s, 1H), 7.46-7.39 (m, 2H),7.32-7.25 (m, 3H), 7.20 (d, J=6.8 Hz, 1H), 7.09-7.06 (m, 1H), 6.99-6.97(m, 2H), 4.53-7.47 (m, 1H), 4.42 (s, 2H), 4.12 (d, J=5.2 Hz, 2H),3.58-3.55 (m, 2H), 3.42-3.32 (m, 2H), 3.20-3.17 (m, 2H), 2.34 (s, 3H)ppm; ESI-MS (m/z): 375.2 [M+1]⁺.

Compound 1171-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-((6-(1-methyl-1H-benzo[d]imidazol-6-yl)pyridin-2-yl)amino)propan-2-ol

Step 1: 6-bromo-N-(oxiran-2-ylmethyl) pyridin-2-amine

To a stirred mixture of 6-bromopyridin-2-amine (1 g, 5.81 mmol) in DMF(15 mL) was added NaH (696 mg, 17.4 mmol), 2-(bromomethyl) oxirane (790mg, 5.81 mmol) at 0° C. The mixture was stirred at 0° C. for 4 hoursthen quenched with water (50 mL), extracted with EA (30 mL×3). Thecombined extracts were washed with brine (20 mL), dried over anhydrousNa₂SO₄ and concentrated. The residue was directly for the next step.LCMS (m/z): 279.1/280.1 [M+H]⁺/[M+2H]⁺

Step 2:1-((6-bromopyridin-2-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a stirred mixture of 6-bromo-N-(oxiran-2-ylmethyl)pyridin-2-amine(1.33 g, 5.81 mmol) in EtOH (10 mL) was added1,2,3,4-tetrahydroisoquinoline (773 mg, 5.81 mmol). The mixture wasstirred at 80° C. for 16 hours. The reaction mixture was concentratedand the residue was purified column to afford the desired compound (140mg). LCMS (m/z): 362.1/363.1 [M+H]⁺/[M+2H]⁺

Step 3:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((6-(1-methyl-1H-benzo[d]imidazol-6-yl)pyridin-2-yl)amino)propan-2-ol

To a stirred mixture of1-((6-bromopyridin-2-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (130 mg, 0.36 mmol) in dioxane:H₂O (15 mL, 2:1) was added1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole(102 mg, 0.396 mmol), Cs₂CO₃ (351 mg, 1.08 mmol) and then Pd(dppf)Cl₂(10 mg). The mixture was degassed with N₂ 4 times and stirred at 25° C.for 16 hours. The reaction mixture was quenched with water (20 mL),extracted with EA (20 mL×3). The combined extracts were washed withbrine (20 mL), dried over anhydrous Na₂SO₄ and concentrated. The residuewas purified by prep-HPLC to afford the title compound (12 mg, 8%).¹HNMR (CH₃OD, 400 MHz) δ: 8.05 (s, 1H), 7.88-7.87 (m, 2H), 7.83-7.80 (m,1H), 7.50 (t, J=8.0, 1H), 7.13-7.11 (m, 4H), 7.01-7.00 (m, 1H), 6.40 (d,J=4.0, 1H), 5.00 (s, 1H), 4.45 (br, 1H), 4.11-4.13 (m, 1H), 3.82-3.61(m, 6H), 3.52-3.45 (m, 1H), 2.91 (s, 3H), 2.81-2.63 (m, 3H). LCMS (m/z):414.2 [M+H]⁺

Compound 1181-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-((4-(1-methyl-1H-benzo[d]imidazol-6-yl)pyridin-2-yl)amino)propan-2-ol

Step 1:1-((4-Bromopyridin-2-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

A mixture of 4-bromo-2-fluoropyridine (100 mg, 0.57 mmol),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (117 mg, 0.57mmol) and TEA (115 mg, 1.14 mmol) in DMF (5 mL) was stirred at 120° C.under microwave heating for 1 h. Water was added to the mixture whichwas extracted with DCM. The combined organic layers were concentrated togive the desired compound as a colorless oil (170 mg, Yield 82%) andused directly in the next step. LCMS (m/z): 362.08, 364.08 (M+1).

Step 2:1-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-((4-(1-methyl-1H-benzo[d]imidazol-6-yl)pyridin-2-yl)amino)propan-2-ol

A mixture of1-((4-bromopyridin-2-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol(170 mg, 0.47 mmol),1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole(177 mg, 0.68 mmol), Pd(dppf)Cl₂ (83 mg, 0.11 mmol) and Na₂CO₃ (151 mg,1.43 mmol) in dioxane/H₂O (5/2 mL) was stirred at 100° C. under N₂ for 2h. Upon completion, water was added to the mixture which was extractedwith DCM. The organic layer was concentrated and purified by pre-HPLC togive the desired title compound (30 mg, yield 16%). ¹H NMR (400 MHz,MeOD): δ 8.41 (s, 1H), 7.90 (d, J=6.8 Hz, 2H), 7.78 (d, J=8.0 Hz, 1H),7.64 (d, J=8.0 Hz, 1H), 7.33-7.19 (m, 4H), 7.04 (d, J=5.6 Hz, 2H), 4.46(s, 2H), 4.35 (br. s, 1H), 3.98 (s, 3H), 3.66-3.53 (m, 4H), 3.33-3.13(m, 4H). LCMS (m/z): 414.2 (M+1).

Compound 1191-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-((4-(1-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)propan-2-ol

Step 1: 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline

To a stirring solution of 1,2,3,4-tetrahydroisoquinoline (13.4 g, 0.1mmol) in acetonitrile (200 mL) was added anhydrous K₂CO₃ (20.7 g, 0.15mol), then 2-(bromomethyl)oxirane (13.7 g, 0.1 mol) in MeCN (20 ml) wasadded drop wise to the reaction. After addition, the solution wasstirred at 25° C. for 4 h until completion of the reaction was observedby LCMS. The solvent was evaporated and the residue purified with columnchromatography to afford the desired product as a colorless oil (11.9 g,80% purity, 50% yield).

Step 2: 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

A sealed vessel was charged with2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (1.5 g, 7.9 mmol)and ammonium in ethanol (50 mL) was then added and the vessel sealed andheated at 80° C. for 2 h. After cooling to room temperature, solventswere evaporated to afford the desired product as a colorless oil (1.2 g,73.7% yield). It was used in the next step without further purification.LCMS (m/z): 207.1 [M+H]⁺

Step 3:1-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-((4-(1-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)propan-2-ol

To a solution of 2,4-dichloropyrimidine (200 mg, 1.36 mmol) in i-PrOH(10 mL) was added TEA (270 mg, 2.72 mmol) and1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl) propan-2-ol (310 mg, 1.5mmol). The reaction mixture was stirred at reflux temperature overnight.The reaction mixture was then concentrated to give a crude mixture of1-((4-chloropyrimidin-2-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-oland1-((2-chloropyrimidin-4-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-olwhich was used for next step without further purification. LCMS (m/z):319.2 [M+H]⁺

To a solution of the crude intermediate mixture in dioxane/H₂O (5 mL)was added1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole(300 mg), Pd(dppf)Cl₂ (67 mg, 0.09 mmol) and Cs₂CO₃ (600 mg, 1.84 mmol).The reaction mixture was heated at 120° C. under microwave condition for40 min. The mixture was concentrated to remove the solvents and theresidue dissolved in ethyl acetate and washed with water. The separatedorganic layer was concentrated and the crude product purified by prepareHPLC to give the title compound (28 mg, 5% overall yield) as a whitesolid.

¹HNMR (CH₃OD, 400 MHz) δ: 8.22-8.20 (m, 2H), 8.15 (s, 1H), 7.96-7.94 (m,1H), 7.62 (d, J=4.6, 1H), 7.15-6.92 (m, 5H), 4.22-4.21 (m, 1H), 3.97 (s,2H), 3.81 (s, 3H), 3.66-3.63 (m, 1H), 3.53-3.48 (m, 1H), 3.12-3.10 (m,2H), 2.99-2.85 (m, 4H). LCMS (m/z): 415.2 [M+H]⁺

Compound 1201-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-((2-(1-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-4-yl)amino)propan-2-ol

The title compound was also isolated from the HPLC purification shownabove yielding1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((2-(1-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-4-yl)amino)propan-2-ol(8 mg, 1.5% (overall)) as a white solid.

¹HNMR (CH₃OD, 400 MHz) δ: 8.32 (s, 1H), 8.17-8.16 (m, 1H), 8.08 (s, 1H),8.03-8.01 (m, 1H), 7.56 (d, J=4.4, 1H), 7.01-6.80 (m, 4H), 6.37-6.36 (m,1H), 4.10 (br, 1H), 3.85-3.64 (m, 6H), 3.50 (br, 1H), 2.83-2.74 (m, 4H),2.72-2.53 (m, 2H). LCMS (m/z): 415.2 [M+H]⁺

Compound 1211-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((6-(1-methyl-1H-benzo[d]imidazol-6-yl)pyrazin-2-yl)amino)propan-2-ol

Step 1:1-((6-chloropyrazin-2-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of 2,6-dichloropyrazine (150 mg, 1 mmol) in 5 mL i-PrOHwas added 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (206mg, 1 mmol), the solution sealed and the mixture heated at 130° C. for 2h. After cooling, the solvent was evaporated and the residue useddirectly in the next step without purification. LCMS (m/z): 319 (M+1).

Step 2:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((6-(1-methyl-1H-benzo[d]imidazol-6-yl)pyrazin-2-yl)amino)propan-2-ol

To a flask containing1-((6-chloropyrazin-2-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol(200 mg, 0.6 mmol) in dioxane:H₂O (3:1) was added1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole(200 mg, 0.8 mmol), Cs₂CO₃ (2 equiv.) and Pd(dppfCl₂) (10 mol %). Themixture was degassed 3 times with N₂ before being heated at 110 deg C.overnight. Upon completion, the solvents were evaporated and residuepurified with prep-HPLC to afford the desired target compound (32 mg,yield 13%). ¹H NMR (400 MHz, MeOD): δ 8.27 (s, 1H), 8.18 (d, J=9.6 Hz,2H), 7.99 (d, J=8.8 Hz, 1H), 7.86 (s, 1H), 7.72 (d, J=8.4 Hz, 1H),7.09-6.99 (m, 3H), 6.87 (d, J=7.6 Hz, 1H), 4.21 (br. s, 1H), 3.86 (s,3H), 3.84-3.54 (m, 4H), 2.89-2.66 (m, 6H). LCMS (m/z): 415.2 (M+1).

Compound 1231-(3-(benzo[d]thiazol-6-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

A mixture of 6-bromobenzo[d]thiazole (200 mg, 0.93 mmol),1-(3,4-dihydroisoquinolin-(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol (420 mg, 1.03 mmol), K₂CO₃ (386 mg, 2.79 mmol) andPd(dppf)Cl₂ (50 mg) in dioxane (8 mL) and H₂O (2 mL) was stirred at 100°C. for 16 h under N₂. The catalyst was filtered and the filtrateconcentrated. The residue was purified by Prep-HPLC to give the titlecompound (101 mg, 26%) as the formate salt. ¹HNMR (CH₃OD, 400 MHz) δ:9.27 (s, 1H), 8.41 (s, 1H), 8.36-8.32 (m, 1H), 8.17-8.11 (m, 1H),7.84-7.82 (m, 1H), 7.49-7.18 (m, 7H), 7.04-7.02 (m, 1H), 4.52-4.48 (m,1H), 4.38 (s, 2H), 4.18-4.14 (m, 2H), 3.56-3.51 (m, 2H), 3.40-3.38 (m,2H), 3.21-3.16 (m, 2H). LCMS (m/z): 417.2 [M+H]⁺

Compound 1251-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(2-(dimethylamino)pyrimidin-5-yl)phenoxy)propan-2-ol

Step 1: 5-bromo-N,N-dimethylpyrimidin-2-amine

A mixture of 5-bromo-2-chloropyrimidine (1 g, 5.18 mmol), dimethylaminehydrochloride (1.26 g, 15.6 mmol) and K₂CO₃ (2.16 g, 15.6 mmol) in EtOH(15 mL) was heated to 120° C. for 16 h. After cooling, the solvent wasevaporated off and the residue was used for next step without furtherpurification. LCMS (m/z): 202.1/203.1 [M+H]⁺/[M+2H]⁺

Step 21-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(2-(dimethylamino)pyrimidin-5-yl)phenoxy)propan-2-ol

A mixture of 5-bromo-N,N-dimethylpyrimidin-2-amine (99 mg, 0,489 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(200 mg, 0.489 mmol), Pd(dppf)Cl₂ (36 mg, 0.049 mmol), K₂CO₃ (202 mg,1.47 mmol) in H₂O-dioxane (1 mL/3 mL) was stirred at 100° C. withmicrowave heating for 15 min. The solvent was removed by concentrationand the crude product was purified by HPLC separation to give the titlecompound as the formate salt (76 mg, 38.5%).

¹HNMR (CH₃OD, 400 MHz) δ: 8.59 (s, 2H), 8.35 (br, 1H), 7.42-7.16 (m,7H), 6.98-6.94 (m, 1H), 4.53-4.46 (m, 3H), 4.18-4.09 (m, 2H), 3.64-3.56(m, 2H), 3.48-3.39 (m, 2H), 3.26-3.17 (m, 8H). LCMS (m/z): 405.2 [M+H]⁺

Compound 126(3′-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)-[1,1′-biphenyl]-3-yl)(morpholino)methanone

Step 1: (3-bromophenyl)(morpholino)methanone

To a solution of 3-bromobenzoic acid (500 mg, 2.49 mmol) in DCM (12 mL)was added morpholine (260 mg, 2.99 mmol), EDCI (1.43 g, 7.47 mmol) andHOBt (1.0 g, 7.47 mmol). The mixture was stirred at room temperature for4 h. The reaction solution was diluted with water and extracted withDCM. The organic layer was concentrated to give the desired(3-bromophenyl)(morpholino) methanone as a crude product which was usedin next step without further purification. LCMS (m/z): 270.1/271.1[M+H]⁺/[M+2H]⁺

Step 2:(3′-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)-[1,1′-biphenyl]-3-yl)(morpholino)methanone

A mixture of (3-bromophenyl)(morpholino)methanone (200 mg, 0.74 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(333 mg, 0.81 mmol), K₂CO₃ (307 mg, 2.22 mmol) and Pd(dppf)Cl₂ (50 mg)in a mixture solution of dioxane (8 mL) and H₂O (2 mL) was stirred at100° C. for 16 h under N₂. The catalyst was filtered and the filtrateconcentrated with the resulting residue purified by Prep-HPLC to givethe title compound as the formate salt (231 mg, 66%). ¹HNMR (CH₃OD, 400MHz) δ: 8.42 (br, 1H), 7.73-7.70 (m, 1H), 7.65 (s, 1H), 7.59-7.50 (m,1H), 7.42-7.35 (m, 2H), 7.28-7.14 (m, 6H), 7.02-6.98 (m, 1H), 4.53-4.49(m, 1H), 4.42-4.34 (m, 2H), 4.14-4.06 (m, 2H), 3.88-3.40 (m, 10H),3.39-3.20 (m, 2H), 3.19-3.06 (m, 2H). LCMS (m/z): 473.2 [M+H]⁺

Compound 1341-((5′-chloro-2′-methoxy-[1,1′-biphenyl]-3-yl)oxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(250 mg, 0.61 mmol) in Dioxane (3 mL) was added2-bromo-4-chloro-1-methoxybenzene (135 mg, 0.61 mmol), 2N K₂CO₃ (1 mL, 2mmol) and PdCl₂(dppf)₂ (20 mg, 0.025 mmol). The reaction mixture washeated at 120° C. under microwave conditions for 30 minutes before beingconcentrated to remove the solvents. The residue was dissolved in ethylacetate, washed with water and the separated organic layer was dried andconcentrated to yield a crude product which was purified by HPLCseparation to give the title compound (102 mg, 39.5%). ¹HNMR (CH₃OD, 400MHz) δ: 7.36-7.28 (m, 7H), 7.10-7.02 (m, 3H), 6.98-6.90 (m, 1H),4.69-4.48 (m, 4H), 4.12-4.03 (m, 2H), 3.78 (s, 3H), 3.70-3.64 (m, 1H),3.52-3.42 (m, 2H), 3.29-3.18 (m, 2H). LCMS (m/z): 424.2 [M+H]⁺

Compound 1411-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenoxy)propan-2-ol

Step 1: 2-Chloro-7-methyl-7H-pyrrolo[2,3-d]pyrimidine

To a solution of NaH (106.6 g, 2.67 mmol) in THF (5 mL) was added2-chloro-7H-pyrrolo[2,3-d]pyrimidine (500 mg, 2.54 mmol) at 0° C. andstirred for 30 minutes at the same temperature. CH₃I (1.5 g, 10.2 mmol)was added at 0° C. and the combined mixture stirred for 3 h at 15° C.After TLC showed the reaction was complete, the mixture was diluted withwater 20 mL) and extracted with ethyl acetate EA (2×20 mL) and thecombined organic layers were washed with brine (30 mL), dried overNa₂SO₄ and concentrated to give the title compound (254 mg, 47.4%) ascolorless oil which was used in next step without further purification.

Step 2:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(7-methyl-7H-pyrrol[2,3-d]pyrimidin-2-yl)phenoxy)propan-2-ol

To a solution of 2-chloro-7-methyl-7H-pyrrolo[2,3-d]pyrimidine (70 mg,0.407 mmol) in dioxane (4 ml) and H₂O (1 mL) was added compound1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(200 mg, 0.4886 mmol), Pd(dppf)Cl₂ (30 mg, 0.041 mmol) and K₂CO₃ (169mg, 1.221 mmol) at 15° C. The reaction mixture was then stirred for 16 hat 100° C. after which, TLC analysis showed the reaction to be complete.The reaction mixture was the filtered and the filtrate concentrated toremove the solvent with the residue purified by prep-HPLC to give theformate salt of the title compound (23 mg, 11.4%) as a white solid. ¹HNMR (400 MHz, CDCl₃): δ 8.99 (s, 1H), 8.10-8.08 (m, 2H), 7.47-7.42 (m,2H), 7.39-7.24 (m, 3H), 7.20 (d, J=6.8 Hz, 1H), 7.10 (dd, J=2.0, 8.0 Hz,1H), 6.66 (d, J=3.6 Hz, 1H), 4.55-4.52 (m, 1H), 4.42 (s, 2H), 4.20-4.17(m, 2H), 3.94 (s, 3H), 3.57-3.54 (m, 2H), 3.45-3.25 (m, 2H), 3.19 (d,J=3.6 Hz, 2H) ppm. LCMS (m/z): 326.1 [M+H]⁺.

Compound 145N-(3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)pyrimidine-5-carboxamide

Step 1: tert-butyl (3-hydroxyphenyl)carbamate

To a solution of compound 3-aminophenol (2.2 g, 0.02 mol) in dioxane (45mL) was added Boc₂O (1034.8 g, 5.2 mmol) under 0° C. The reactionmixture was stirred at 0° C. for 3 h then concentrated with the residuethen dissolved in ethyl acetate. This organic layer was washed with 1MNaOH (100 ml) solution, water then dried over sodium sulfate, filteredand concentrated to yield a crude product which was used in next stepwithout further purification.

Step 2: tert-butyl (3-(oxiran-2-ylmethoxy)phenyl)carbamate

To a solution of tert-butyl (3-hydroxyphenyl)carbamate (1 g, 4.79 mmol)in DMF (15 mL) was added NaH (0.126 g, 5.26 mmol) at 0° C. Afterstirring for 30 min, 2-(bromomethyl)oxirane (0.716 g, 5.26 mmol) wasadded and the mixture stirred at 0° C. for 1 h before quenching with theaddition of MeOH. After evaporation of this mixture, the resultingresidue was dissolved in ethyl acetate and washed with water, dried andwith sodium sulfate and concentrated with the resulting crude productused for next step without further purification.

Step 3: tert-butyl(3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)carbamate

To a solution of tert-butyl (3-(oxiran-2-ylmethoxy)phenyl)carbamate 1.5g, 5.66 mmol) in EtOH (30 mL) was added 1,2,3,4-tetrahydroisoquinoline0.828 g, 6.23 mmol). The reaction mixture was heated at 100° C. for 4 h.The solvent was then removed by concentration and the residue dissolvedin ethyl acetate, washed with water and the separated organic layerdried over sodium sulfate and concentrated to obtain a crude product.This product was used in next step without further purification.

Step 4:1-(3-aminophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of tert-butyl(3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)carbamate(1.2 g, 3.01 mmol) in ethyl acetate (30 mL) was added HCl/ethyl acetate(20 mL). The reaction mixture was stirred at room temperature for 2 h.and the concentrated with the residue used in the next step withoutfurther purification.

Step 5:N-(3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)pyrimidine-5-carboxamide

To a solution of pyrimidine-5-carboxylic acid (41 mg, 0.33 mmol) in DCM(10 mL) was added HATU (120.54 g, 0.33 mmol) and (Et)₃N (64.9 g, 0.642mmol). After stirred at room temperature for 30 min,1-(3-aminophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (100mg 0.32 mmol) was added and the combined reaction mixture was stirredfor 1 h. The residue was diluted with DCM (100 mL) and washed by water(30 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered andconcentrated to obtain the crude product which was purified bypreparative HPLC separation to give the desired product as the formatesalt (37.5 g, 38.5%). ¹H NMR (400 MHz, CDCl₃): δ 10.70 (s, 1H), 9.30 (d,J=1.6 Hz, 1H), 8.95 (d, J=2.8 Hz, 1H), 8.82 (dd, J=1.6, 2.4 Hz, 1H),8.17 (s, 1H), 7.63-7.62 (m, 1H), 7.50 (d, J=8 Hz, 1H), 7.29-7.25 (m,1H), 7.11-7.05 (m, 4H), 6.76-6.74 (m, 1H), 4.12-4.10 (m, 1H), 4.06-4.02(m, 1H), 3.94-3.89 (m, 1H), 3.68 (s, 1H), 2.82-2.65 (m, 4H), 2.61-2.56(m, 2H) ppm. LCMS (m/z): 305.2 [M+H]⁺.

Compound 1566-(3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)-2H-benzo[b][1,4]oxazin-3(4H)-one

To a solution of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(200 mg, 0.49 mmol) in Dioxane (3 mL) was added6-bromo-2H-benzo[b][1,4]oxazin-3(4H)-one (112 mg, 0.49 mmol), 2N K₂CO₃(1 mL, 2 mmol) and PdCl₂(dppf)₂ (20 mg, 0.025 mmol). The reactionmixture was heated at 120° C. under microwave conditions for 30 min. Themixture was then concentrated to remove the solvents and the residue wasdissolved in ethyl acetate, washed with water with the separated organiclayers dried before concentrating again. The resulting residue was thenpurified by prep-HPLC to afford the title compound as the formate salt(62 mg, 29.4%). ¹H NMR (400 MHz, MeOD): δ 8.40 (brs, 1H), 7.36-7.12 (m,8H), 7.00 (d, J=8.4 Hz, 1H), 6.95-6.93 (m, 1H), 4.59 (s, 2H), 4.51-4.48(m, 1H), 4.42 (s, 1H), 4.11-4.10 (m, 2H), 3.57-3.54 (m, 2H), 3.42-3.38(m, 2H), 3.20-3.16 (m, 2H) ppm; ESI-MS (m/z): 431.2 [M+1]⁺.

Compound 1671-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((2′-phenoxy-[1,1′-biphenyl]-3-yl)oxy)propan-2-ol

Step 1: Bromo-2-phenoxybenzene

To a solution of 2-bromophenol (1 g, 5.84 mmol), phenylboronic acid(1.42 g, 11.69 mmol) and TEA (2.95 g, 29.2 mmol) in DCM (30 mL) wasadded Cu(OAc)₂ (1.05 g, 5.84 mmol) and 4 Å molecular sieves (500 mg).The reaction mixture was stirred at room temperature over air with a drytube attached for 16 h. The mixture was filtered and the filtrate waswashed by water (50 mL) and brine (30 mL). The organic layer was driedover anhydrous Na₂SO₄, filtered and concentrated with the residuepurified by column chromatography to give 612 mg of the desired product.

Step 2:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((2′-phenoxy-[1,1′-biphenyl]-3-yl)oxy)propan-2-ol

A mixture of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(200 mg, 0.489 mmol), 1-bromo-2-phenoxybenzene (120 mg, 0.489 mmol),Pd(dppf)Cl₂ (36 mg, 0.049 mmol), K₂CO₃ (202 mg, 1.47 mmol) inH₂O-dioxane (1 mL/3 mL) was stirred at 100° C. under microwave heatingfor 15 min. The solvent was removed and the crude product purified byprep-HPLC to afford the title compound as the formate salt (33 mg, 15%).¹H NMR (400 MHz, MeOD): δ 8.46 (s, 1H), 7.49-7.22 (m, 9H), 7.17-7.11 (m,3H), 7.04-6.98 (m, 2H), 6.92-6.90 (m, 1H), 6.87-6.84 (m, 1H), 4.43-4.37(m, 1H), 4.28 (s, 2H), 3.98 (d, J=5.2 Hz, 2H), 3.41 (t, J=6.4 Hz, 2H),3.25-3.13 (m, 4H) ppm; ESI-MS (m/z): 452.3 [M+1]⁺.

Compound 1761-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-methyl-1H-pyrrolo[2,3-b]pyridin-6-yl)phenoxy)propan-2-ol

Step 1: 6-Bromo-1-methyl-1H-pyrrolo[2,3-b]pyridine

To a solution of 6-bromo-1H-pyrrolo[2,3-b]pyridine (500 mg, 2.55 mmol)in DMF (10 mL) was added NaH (306 mg, 7.65 mmol, m %/60%). The mixturewas stirred at room temperature for 1 h and then MeI (398 mg, 2.8 mmol)was added. The reaction mixture was then stirred at room temperature for4 h after which, TLC showed the completion of the reaction. The reactionmixture was quenched by addition of water and then extracted with EtOAc.Combined organic layers were concentrated and the residue was purifiedby Prep-TLC (PE:EA=5:1) to give the desired product (200 mg) which wasused directly in the next step.

Step 2:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-methyl-1H-pyrrolo[2,3-b]pyridin-6-yl)phenoxy)propan-2-ol

A mixture of 6-bromo-1-methyl-1H-pyrrolo[2,3-b]pyridine (200 mg, 0.95mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(426 mg, 1.04 mmol), K₂CO₃ (394 mg, 2.85 mmol) and Pd(dppf)Cl₂ (50 mg)in a solution of dioxane (8 mL) and H₂O (2 mL) was stirred at 100° C.for 16 h under N₂. The catalyst was filtered and the filtrateconcentrated with the resulting residue purified by prep-HPLC to affordthe title compound as the formate salt (139 mg, 35.4%). ¹H NMR (400 MHz,MeOD): δ 8.43 (s, 1H), 7.99 (d, J=8.4 Hz, 1H), 7.77-7.57 (m, 3H),7.43-7.36 (m, 2H), 7.31-7.18 (m, 4H), 7.03-7.01 (m, 1H), 6.49 (d, J=3.6Hz, 1H), 4.52 (dd, J=8.8, 4 Hz, 1H), 4.41 (s, 2H), 4.19-4.14 (m, 2H),3.93 (s, 3H), 3.57-3.54 (m, 2H), 3.43-3.33 (m, 2H), 3.19-3.16 (m, 2H)ppm; ESI-MS (m/z): 414.3 [M+1]⁺.

Compound 1871-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1,3-dimethyl-1H-indol-6-yl)phenoxy)propan-2-ol

Step 1: 6-Bromo-1H-indole-3-carbaldehyde

POCl₃ (980 mg, 6.4 mmol) was added dropwise to DMF (3 mL) cooled in anice bath. The mixture was stirred at 0° C. for 30 min before a solutionof 6-bromo-1H-indole (1.0 g, 5.1 mmol) in DMF (7 mL) was slowly added at0° C. The mixture was stirred at room temperature for 3 h before beingpoured into water and neutralized with 1N NaOH. The crude product wascollected by filtration and used in next step without furtherpurification. LCMS (m/z): 224.1 [M+H]⁺.

Step 2: 6-Bromo-3-methyl-1H-indole

To a solution of 6-bromo-1H-indole-3-carbaldehyde (300 mg, 1.34 mmol) inTHF (10 mL) was added LiAlH₄ (100 mg, 2.63 mmol) at 0° C. The mixturewas heated to 70° C. and stirred for 4 h at this temperature. Uponcooling, the reaction solution was quenched with 40% of NaOH andfiltered and the filtrate concentrated to give crude product which wasused in next step without further purification. LCMS (m/z): 210.1[M+H]⁺.

Step 3: 6-Bromo-1,3-dimethyl-1H-indole

To a solution of 6-bromo-3-methyl-1H-indole (200 mg, 0.95 mmol) in DMF(8 mL) was added NaH (114 mg, 2.85 mmol, m %/60%). The mixture wasstirred at room temperature for 1 h before the addition of MeI (162 mg,1.14 mmol). The system was stirred at room temperature for 4 h then themixture was quenched with water and extracted with EtOAc. The organiclayer was concentrated to give crude product which was used in next stepwithout further purification. LCMS (m/z): 224.1 [M+H]⁺.

Step 4:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1,3-dimethyl-1H-indol-6-yl)phenoxy)propan-2-ol

A mixture of 6-bromo-1,3-dimethyl-1H-indole (200 mg, 0.89 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(401 mg, 0.98 mmol), K₂CO₃ (369 mg, 2.67 mmol) and Pd(dppf)Cl₂ (50 mg)in dioxane (8 mL) and H₂O (2 mL) was stirred at 100° C. for 16 h underN₂. The catalyst was filtered and the filtrate was concentrated. Theresidue was purified by prep-HPLC to afford the title compound (38 mg,Yield 10%). ¹H NMR (400 MHz, MeOD): δ 8.42 (s, 1H), 7.52-7.50 (m, 2H),7.37-7.25 (m, 7H), 7.16-7.14 (m, 1H), 6.93-6.89 (m, 2H), 4.48-4.45 (m,1H), 4.34 (s, 2H), 4.11-4.09 (m, 2H), 3.76 (s, 3H), 3.50-3.47 (m, 2H),3.36-3.31 (m, 2H), 3.31-3.14 (m, 2H), 2.30 (s, 3H) ppm; ESI-MS (m/z):427.3 [M+1]⁺.

Compound 1891-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(3-methyl-3H-imidazo[4,5-b]pyridin-5-yl)phenoxy)propan-2-ol

Step 1: 6-Chloro-N-methyl-3-nitropyridin-2-amine

To a solution of 2,6-dichloro-3-nitropyridine (1 g, 7.3 mmol) in EtOH(10 mL) was added Na₂CO₃ (1.3 g, 12.3 mmol) and MeNH₂ at 18° C. Themixture was stirred for 16 hours at 18° C. after which TLC analysisshowed the reaction was complete. The reaction mixture was thenconcentrated to get the crude material which was treated with water (50mL) and extracted with ethyl acetate. The combined organic layers werewashed with brine (30 mL), dried over Na₂SO₄ and concentrated to getyield the crude product which was purified by crystallized with ethanolto give the desired product (780 mg, 55.7%) as a yellow solid. LCMS(m/z): 188.1 [M+H]⁺.

Step 2: 6-Chloro-N₂-methylpyridine-2,3-diamine

To a solution of 6-chloro-N-methyl-3-nitropyridin-2-amine (630 mg, 3.36mmol) in MeOH (10 ml) and water (10 ml) was added Fe (940.8 g, 16.8mmol) and NH₄Cl (898.8 g, 168 mmol) at 20° C. The mixture was stirredfor 3 hours under reflux temperature until TLC (PE:EA=3:1) showed thatthe reaction was complete. The mixture was filtered, the filtrateconcentrated and residue treated with water (10 ml) and extracted withethyl acetate (2×20 ml). The combined organic layers were washed withbrine (30 ml), dried over Na₂SO₄ and concentrated to give the desiredproduct (316 mg, 59.7%) as a yellow solid which was used in next stepwithout further purification. LCMS (m/z): 158.1 [M+H]⁺.

Step 3: 5-Chloro-3-methyl-3H-imidazo[4,5-b]pyridine

To a solution of 6-chloro-N₂-methylpyridine-2,3-diamine (300 mg, 1.9mmol) in HC(OMe)₃ (10 mL) was added TsOH (10 mg) at 22° C. The mixturewas stirred for 3 h under reflux until TLC (PE:EA=3:1) showed that thereaction was complete. The mixture was concentrated to get the crudematerial which was purified by using column chromatography on silica gelto give the desired product (261 mg, 81.6%) as an oil. LCMS (m/z): 168.1[M+H]⁺.

Step 4:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(3-methyl-3H-imidazo[4,5-b]pyridin-5-yl)phenoxy)propan-2-ol

To a solution of 5-chloro-3-methyl-3H-imidazo[4,5-b]pyridine (55 mg,0.3257 mmol) in dioxane (4 ml) and H₂O (1 mL) was added1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(200 mg, 0.4886 mmol), Pd(dppf)Cl₂ (30 mg, 0.04 mmol) and K₂CO₃ (135 mg,0.9771 mmol) at 21° C. The mixture was stirred for 16 h at 100° C. atwhich point TLC (PE:EA=1:1) showed that the reaction was complete. Themixture was concentrated to get the crude which was purified byprep-HPLC to afford the title compound as the formate salt (32 mg, Yield23.7%). ¹H NMR (400 MHz, MeOD): δ 8.37 (s, 1H), 8.12-8.10 (m, 1H),7.86-7.72 (m, 3H), 7.44 (t, J=8 Hz, 1H), 7.28-7.17 (m, 4H), 7.08-7.06(m, 1H), 4.51-4.48 (m, 1H), 4.34 (s, 2H), 4.19-4.18 (m, 2H), 4.00 (s,3H), 3.49-3.46 (m, 2H), 3.36-3.29 (m, 2H), 3.28-3.16 (m, 2H) ppm; ESI-MS(m/z): 415.3 [M+1]⁺.

Compound 2081-(3-(1-(azetidin-3-yl)-1H-benzo[d]imidazol-6-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

Step 1: tert-butyl3-((5-bromo-2-nitrophenyl)amino)azetidine-1-carboxylate

To a solution of 4-bromo-2-fluoro-1-nitrobenzene (500 mg, 2.27 mmol) andtert-butyl 3-aminoazetidine-1-carboxylate (390 mg, 2.27 mmol) in DMSO (5mL) was added DIPEA (351 mg, 2.72 mmol). The solution was stirred insealed tube at room temperature for 16 h after which time, LCMS showedthe completion of the reaction. The solution was diluted with water (100mL) and DCM (100 mL) and the organic layer washed with water (30 mL×2)and brine (30 mL) and dried over anhydrous Na₂SO₄, then filtered andconcentrated to give desired crude product (660 mg, Yield 80%). LCMS(m/z): 372.1 (M+1) which was used in the next step without furtherpurification.

Step 2: tert-butyl3-((2-amino-5-bromophenyl)amino)azetidine-1-carboxylate

To a solution of crude tert-butyl3-((5-bromo-2-nitrophenyl)amino)azetidine-1-carboxylate (550 mg, 1.5mmol) in EtOH (10 mL) was added Fe powder (1.27 g, 22.7 mmol) and NH₄Cl(1.20 g, 0.227 mmol, in 10 ml water). After stirring for 4 h atrefluxing temperature, the solution cooled and filtered through a Celitepad and the filtrate concentrated. The residue used directly for nextstep (400 mg, Yield 78%) without further purification. LCMS (m/z): 342.1(M+1).

Step 3: tert-butyl3-(6-bromo-1H-benzo[d]imidazol-1-yl)azetidine-1-carboxylate

To a solution of tert-butyl3-((2-amino-5-bromophenyl)amino)azetidine-1-carboxylate (320 mg, 0.938mmol), HC(OCH₃)₃ (5 mL) in DMF (10 mL) was added concentrated HCl (twodrops). The reaction mixture was stirred at room temperature for 16 hthen concentrated and diluted with DCM (50 mL) and washed with saturatedNaHCO₃ solution (25 mL). The organic layer was washed with water (50 mL)and dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated to yield a crude desired product (250 mg, Yield 80%) whichwas used in the next step without further purification. LCMS (m/z):352.1 (M+1).

Step 4: 1-(Azetidin-3-yl)-6-bromo-1H-benzo[d]imidazole

To a solution of tert-butyl3-(6-bromo-1H-benzo[d]imidazol-1-yl)azetidine-1-carboxylate (crude) inDCM (9 mL) was added TFA (3 mL). After stirred for 2 h, the solution wasconcentrated to give desired product and used directly in the next step.

Step 5:1-(3-(1-(azetidin-3-yl)-1H-benzo[d]imidazol-6-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

A mixture of 1-(azetidin-3-yl)-6-bromo-1H-benzo[d]imidazole (100 mg,0.389 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy) propan-2-ol (162 mg, 0.389 mmol), Pd(dppf)Cl₂ (30 mg, 0.039mmol) and K₂CO₃ (265 mg, 1.17 mmol) in H₂O-dioxane (1.5 mL/4.5 mL) wasstirred at 100° C. under microwave heating for 15 minutes. The solventwas removed by concentration and the crude product purified by HPLCseparation to yield the target compound as the formate salt (40 mg,Yield 23%). ¹H NMR (400 MHz, MeOD) δ 8.55 (s, 1H), 7.92 (s, 1H), 7.79(d, J=8.4 Hz, 1H), 7.62 (dd, J=8.4 Hz, 1H), 7.43 (dd, J=8.0 Hz, 1H),7.34-7.17 (m, 6H), 7.01 (dd, J=8.0 Hz, J₂=1.2 Hz, 1H), 5.85-5.86 (m,1H), 4.79 (t, J=8.0 Hz, 2H), 4.66 (t, J=8.0 Hz, 2H), 4.39 (s, 2H), 4.17(d, J=4.8 Hz, 1H), 3.55 (t, J=4.8 Hz, 2H), 3.42-3.12 (m, 4H). LCMS(m/z): 452.2 (M+1).

Compound 2091-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-(pyrrolidin-3-yl)-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

Step 1: tert-butyl3-((5-bromo-2-nitrophenyl)amino)pyrrolidine-1-carboxylate

To a solution of 4-bromo-2-fluoro-1-nitrobenzene (500 mg, 2.27 mmol) andtert-butyl 3-aminopyrrolidine-1-carboxylate (390 mg, 2.27 mmol) in DMSO(5 mL) was added DIPEA (351 mg, 2.72 mmol) and the solution stirred in asealed bottle at room temperature for 16 h. After LCMS analysisindicated completion of the reaction, the solution was diluted withwater (100 mL) and DCM (100 mL). The organic layer was washed by water(30 mL×2) and brine (30 mL) and dried over anhydrous Na₂SO₄ before beingfiltered and concentrated to yield the desired product (670 mg Yield80%). The material was used in the next step without furtherpurification. LCMS (m/z): 386.1 (M+1).

Step 2: tert-butyl3-((2-amino-5-bromophenyl)amino)pyrrolidine-1-carboxylate

To a solution of tert-butyl3-((5-bromo-2-nitrophenyl)amino)pyrrolidine-1-carboxylate (crude) (550mg, 1.5 mmol) in EtOH (10 mL) was added Fe powder (1.27 g, 22.7 mmol)and NH₄Cl (1.20 g, 0.227 mmol, in 10 ml water). After stirring for 4 hat refluxing temperature, the reaction mixture was filtered over aCelite pad and the filtrate concentrated. The residue used for next stepwithout further purification (410 mg, Yield 76%). LCMS (m/z): 356.1(M+1).

Step 3: tert-butyl3-(6-bromo-1H-benzo[d]imidazol-1-yl)pyrrolidine-1-carboxylate

To a solution of tert-butyl3-((2-amino-5-bromophenyl)amino)pyrrolidine-1-carboxylate (320 mg, 0.938mmol), HC(OCH₃)₃ (5 mL) in DMF (10 mL) was added concentrated HCl (twodrops). The mixture was then stirred at room temperature for 16 h beforeconcentration. The residue was then diluted with DCM (50 mL) and washedwith saturated NaHCO₃ solution (25 mL), water (50 mL), dried overanhydrous Na₂SO₄ and filtered. The filtrate was concentrated to give thecrude desired product (250 mg, Yield 80%) which was used on the nextstep without further purification. LCMS (m/z): 366.1 (M+1).

Step 4: 6-bromo-1-(pyrrolidin-3-yl)-1H-benzo[d]imidazole

To a solution of tert-butyl3-(6-bromo-1H-benzo[d]imidazol-1-yl)pyrrolidine-1-carboxylate (crude) inDCM (9 mL) was added TFA (3 mL). After stirring for 2 h, the solutionwas concentrated to give the desired crude product which was useddirectly in the next step.

Step 5:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-(pyrrolidin-3-yl)-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

A mixture of 6-bromo-1-(pyrrolidin-3-yl)-1H-benzo[d]imidazole (100 mg,0.389 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy) propan-2-ol (162 mg, 0.389 mmol), Pd(dppf)Cl₂ (30 mg, 0.039mmol) and K₂CO₃ (265 mg, 1.17 mmol) in H₂O-dioxane (1.5 mL/4.5 mL) wasstirred at 100° C. over microwave for 15 min. The solvent was thenremoved by concentration and the crude product purified by HPLCseparation to yield the target title compound as the formate salt (40mg, Yield 23%).

¹H NMR (400 MHz, MeOD) δ 8.42 (s, 1H), 7.88 (s, 1H), 7.78 (d, J=8.4 Hz,1H), 7.62 (dd, J=8.4 Hz, 1H), 7.42 (dd, J=7.6 Hz, 1H), 7.34-7.17 (m,6H), 7.01 (dd, J=8.0 Hz, J₂=1.2 Hz, 1H), 5.49-5.46 (m, 1H), 4.51-4.48(m, 1H), 4.33 (s, 2H), 4.17 (d, J=4.8 Hz, 1H), 3.98-3.93 (m, 1H),3.74-3.46 (m, 5H), 3.36-3.15 (m, 4H), 2.76-2.66 (m, 2H). LCMS (m/z):469.32 (M+1).

Compound 2101-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(2-methyl-2H-pyrazolo[3,4-b]pyridin-6-yl)phenoxy)propan-2-ol

Step 1: (2,6-dichloropyridin-3-yl)methanol

To a solution of 2,6-dichloronicotinic acid (1 g, 5.2 mmol) in THF (10mL) was added NaBH₄ (591 mg, 15.6 mmol) at 0° C. The mixture was stirredfor 30 min and then BF₃.OEt₂ (2.2 g, 15.6 mmol) was added drop wise at0° C. After addition was complete, the mixture was stirred at roomtemperature for 10 hr, until the reaction was completed. The reactionmixture was quenched by the addition of saturated NH₄Cl solution (50 mL)and extracted with ethyl acetate (3×30 mL). The combined organic layerswere washed with brine (30 mL), dried over Na₂SO₄ and concentrated togive the desired product as a white solid which was used in next stepwithout further purification. (820 mg, Yield 80%).

Step 2: 2,6-dichloronicotinaldehyde

To a solution of (2,6-dichloropyridin-3-yl)methanol (1.0 g, 5.62 mmol)in CH₂Cl₂ (10 ml) was added Dess-Martin reagent (4.8 g, 11.24 mmol) at26° C. After addition the mixture was stirred at room temperature for 2h. Once the reaction was complete, the mixture was then quenched byadding 5% aqueous Na₂S₂O₃ and stirred for 30 min. The resulting mixturewas extracted with CH₂Cl₂ (2×30 ml). The combined organic layers werewashed with saturated Na₂S₂O₃ solution (50 ml), brine (30 ml), driedover Na₂SO₄ and concentrated to give the title compound which was usedin next step without further purification. (800 mg, Yield 80%). ¹H NMR(400 MHz, CDCl₃): 10.38 (s, 1H), 8.19 (d, J=8.0 Hz, 1H), 7.44 (d, J=8.0Hz, 1H).

Step 3: 6-chloro-2-methyl-2H-pyrazolo[3,4-b]pyridine

To a solution of 2,6-dichloronicotinaldehyde (600 mg, 3.41 mmol) in THF(5 mL) was added CH₃NHNH₂ (480 mg, 4.1 mmol) in a seal tube at 26° C.The mixture was stirred for 16 h at 120° C. until the reaction wascomplete. The mixture was then treated with water (20 ml) and extractedwith ethyl acetate (3×20 ml) and the combined organic layers washed withbrine (20 ml), dried over Na₂SO₄ and concentrated to get the crudematerial, which was purified by column chromatography to yield thedesired compound as a yellow solid (380 mg, 72.4%) (420 mg, Yield 75%).

¹H NMR (400 MHz, MeOD): 8.23 (d, J=8.4 Hz, 1H), 7.52 (s, 1H), 7.36 (d,J=8.4 Hz, 1H), 2.98 (s, 3H).

Step 4:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(2-methyl-2H-pyrazolo[3,4-b]pyridin-6-yl)phenoxy)propan-2-ol

To a solution of 6-chloro-2-methyl-2H-pyrazolo[3,4-b]pyridine (220 mg,1.313 mmol) in dioxane (8 ml) and H₂O (2 mL) was added1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(800 mg, 1.969 mmol), Pd(dppf)Cl₂ (16 mg, 0.131 mmol) and K₂CO₃ (544 mg,3.939 mmol) at 27° C. The reaction mixture was then stirred for 16 h at100° C. until the reaction was shown to be complete by TLC analysis. Themixture was then concentrated to a crude material which was purified byprep-HPLC separation to give the desired product1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(2-methyl-2H-pyrazolo[3,4-b]pyridin-6-yl)phenoxy)propan-2-olas a white solid (13 mg, 2.4%). ¹H NMR (400 MHz, MeOD): 8.13 (d, J=8.4Hz, 1H), 8.02 (s, 1H), 7.80 (s 1H), 7.74 (d, J=8.0 Hz, 1H), 7.63 (d,J=8.4 Hz, 1H), 7.46 (dd, J₁=J₂=8.0 Hz, 1H), 7.29 (s, 1H), 7.20-7.15 (m,3H), 7.07-7.05 (m, 2H), 4.35-4.32 (m, 1H), 4.24 (s, 3H), 4.21-4.15 (m,2H), 3.98 (d, J=6.8 Hz, 2H), 3.80 (d, J=6.8 Hz, 2H), 3.08-2.86 (m, 6H).LCMS (m/z): 415.2 (M+1).

Compound 2161-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-methyl-4,5-dihydro-1H-pyrazolo[3,4-c]pyridin-6(7H)-yl)phenoxy)propan-2-ol

Step 1: tert-butyl4-((dimethylamino)methylene)-3-oxopiperidine-1-carboxylate

To a stirred mixture of tert-butyl 3-oxopiperidine-1-carboxylate (1 g, 5mmol) was added DMF-DMA (2 mL). The mixture was stirred at 100° C. for 2hours. The mixture was concentrated by vacuum and the residue wasdirectly for the next step (1.20 g, yield: 94%).

Step 2: tert-butyl4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate

To a stirred mixture of tert-butyl4-((dimethylamino)methylene)-3-oxopiperidine-1-carboxylate (1.0 g, 4mmol) in EtOH (10 mL) was added hydrazine hydrate (376 mg, 10 mmol). Themixture was stirred at 80° C. for 16 hours until the reaction was shownto be complete by LCMS. The mixture was then concentrated and theresidue was extracted with EA (3×30 mL) from water (30 mL). The combinedorganic extracts were washed with brine (20 mL), dried over anhydrousNa₂SO₄ and concentrated. The residue was directly for the next step (810mg, 90% yield). LCMS: 224.1 (M+1), 114.1 (M+1-100), 168.1 (M+1-56).

Step 3: tert-butyl1-methyl-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate

To a stirred mixture of tert-butyl4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate (700 mg, 3.14mmol) in DMF (5 mL) was added NaH (251 mg, 6.28 mmol) at 0° C. and thenMeI (669, 4.71 mmol). The mixture was then stirred at 25° C. for 4 hoursafter which the reaction was quenched by addition of water (50 mL) andextracted with ethyl acetate (3×50 mL). The combined extracts werewashed with brine (20 mL), dried over anhydrous Na₂SO₄ and concentratedto yield a residue which was used directly in the next step. ¹H NMR (400MHz, MeOD): δ 7.16 (s, 1H), 3.77 (s, 2H), 3.66 (s, 3H), 3.06 (t, J=6.0Hz, 2H), 2.56 (t, J=6.0 Hz, 2H).

Step 4: 1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine

To a stirred mixture of tert-butyl1-methyl-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate (744mg, 3.14 mmol) in EA (2 mL) was added HCl/EA (3 N, 4 mL). The mixturewas stirred at 25° C. for 16 hours. The reaction mixture was thenconcentrated and the residue purified by Prep-HPLC and SFC to afford thetarget compound as colorless oil which was used without furtherpurification in the next step. (120 mg; yield 28%). LCMS (m/z): 138.10(M+1).

Step 5:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-methyl-4,5-dihydro-1H-pyrazolo[3,4-c]pyridin-6(7H)-yl)phenoxy)propan-2-ol

To a stirred mixture of1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine (30 mg, 0.219mmol) in dioxane:H₂O (6 mL, 2:1) was added1-(3-bromophenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (119mg, 0.328 mmol), Cs₂CO₃ (213 mg, 0.657 mmol) and then Ruphos (2 mg) andPd(Ruphos) (5 mg). The mixture was degassed by N₂ 4 times and stirred at100° C. for 16 hours. The reaction mixture was then quenched with water(20 mL), extracted with ethyl acetate (3×20 mL). The combined extractswere washed with brine (20 mL), dried over anhydrous Na₂SO₄ andconcentrated to yield a residue which was purified by prep-HPLC toafford1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-methyl-4,5-dihydro-1H-pyrazolo[3,4-c]pyridin-6(7H)-yl)phenoxy)propan-2-ol(17 mg, Yield 18.6%). ¹H NMR (400 MHz, MeOD) δ=7.32 (s, 1H), 7.19-7.08(m, 4H), 7.04 (d, J=5.8 Hz, 1H), 6.70-6.61 (m, 2H), 6.50-6.43 (m, 1H),4.29-4.22 (m, 1H), 4.20 (s, 2H), 4.09-4.01 (m, 1H), 4.01-3.92 (m, 1H),3.76 (s, 2H), 3.73 (s, 3H), 3.61 (t, J=5.8 Hz, 2H), 2.98-2.84 (m, 4H),2.82-2.67 (m, 4H). LCMS (m/z): 419.2 (M+1).

Compound 2181-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-(1-methylazetidin-3-yl)-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

Step 1: 6-bromo-1-(1-methylazetidin-3-yl)-1H-benzo[d]imidazole

To a solution of 1-(azetidin-3-yl)-6-bromo-1H-benzo[d]imidazole (100 mg,0.396 mmol) in MeOH (10 mL) was added HCHO (300 mg, 3.96 mmol) and AcOH(two drops). The reaction mixture was stirred at room temperature for 30min. NaCNBH₃ (29.7 g, 0.476 mmol) was then added and the reactionmixture stirred at room temperature for 2 h. The solvents were thenremoved by concentration and the residue was used for next step withoutpurification. ESI-MS (m/z): 266.1 [M+1]

Step 2:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-(1-methylazetidin-3-yl)-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

A mixture of 6-bromo-1-(1-methylazetidin-3-yl)-1H-benzo[d]imidazole (105mg, 0.389 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(162 mg, 0.389 mmol), Pd(dppf)Cl₂ (30 mg, 0.039 mmol) and K₂CO₃ (265 mg,1.17 mmol) in H₂O-dioxane (1.5 mL/4.5 mL) was stirred at 100° C. undermicrowave heating for 15 min. The solvent was then removed byconcentration and the residue purified by prep-HPLC to afford the titlecompound as a formate salt (14.3 g, yield: 7.7%). ¹H NMR (400 MHz,MeOD): δ 8.57 (s, 1H), 8.43 (s, 1H), 7.84-7.77 (m, 2H), 7.62-7.60 (m,1H), 7.44-7.40 (m, 1H), 7.35-7.25 (m, 5H), 7.21 (d, J=7.2 Hz, 1H), 7.00(dd, J=8, 1.6 Hz, 1H), 5.53-5.49 (m, 1H), 4.53 (dd, J=9.2, 4 Hz, 1H),4.46 (s, 2H), 4.40-4.36 (m, 2H), 4.25-4.16 (m, 4H), 3.60 (t, J=6.4 Hz,2H), 3.47-3.43 (m, 2H), 3.33-3.21 (m, 2H), 2.81 (s, 3H) ppm; ESI-MS(m/z): 469.3 [M+1]⁺.

Compound 2231-(3-(1H-benzo[d]imidazol-4-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

Step 1: 3-bromobenzene-1,2-diamine

A mixture of 3-bromo-2-nitroaniline (2 g, 9.2 mmol), Fe powder (1.54 g,27.6 mmol) and NH₄Cl (4.8 g, 52 mmol) in EtOH and H₂O was stirred underreflux for 1 h. The mixture was cooled to room temperature and filtered.The filtrate was diluted water extracted with ethyl acetate and thecombined organic layers were dried and concentrated in vacuum to givethe crude desired product (1.7 g, Yield 85%). This crude was used in thenext step without further purification.

Step 2: 4-bromo-1H-benzo[d]imidazole

A mixture of 3-bromobenzene-1,2-diamine (2 g, 10.7 mmol) in HCOOH wasstirred at 100° C. for 1 h. The mixture was cooled and concentrated invacuo and diluted with ethyl acetate and aq. NaHCO₃. The mixture wasextracted further with ethyl acetate and the combined organic layersdried and concentrated to give the crude product which was purified bycolumn chromatography to obtain the desired product (1.65 g, Yield 80%).LCMS (m/z): 197.0 (M+1).

Step 3:4-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole

To a solution of 4-bromo-1H-benzo[d]imidazole (1.1 g, 5.6 mmol) in THFwas added NaH (448 mg, 11.2 mmol) at 0° C. After stirring at for 30minutes at room temperature 0° C., SEM-Cl (1.4 g, 8.4 mmol) was added tothe mixture at 0° C. The mixture was then stirred at room temperaturefor 16 h, at which time TLC showed the completion of the reaction. Thereaction was quenched by addition of aq. NH₄Cl and the mixture extractedwith ethyl acetate with the combined organic layers were dried overanhydrous Na₂SO₄ and concentrated in vacuum to give the desired productand used in the next step without further purification (1.65 g, Yield90%). LCMS (m/z): 229.0 (M+1)

Step 4:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-4-yl)phenoxy)propan-2-ol

A mixture of4-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole (200mg, 0.611 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(375 mg, 0.916 mmol), Pd(dppf)Cl₂ (89 mg, 0.12 mmol) and Na₂CO₃ (130 mg,1.22 mmol) in dioxane and H₂O was stirred under N₂ atmosphere at 100° C.for 3 h. The mixture was extracted with DCM and the combined organiclayers dried over Na₂SO₄ and concentrated in vacuum to give the crudeproduct (265 mg, Yield 82%) which was used without further purification.

Step 5:1-(3-(1H-benzo[d]imidazol-4-yl)phenoxy)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

A mixture of 1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-((2-(trimethylsilyl) ethoxy)methyl)-1H-benzo[d]imidazol-4-yl) phenoxy) propan-2-ol (265 mg, 0.5mmol) in EtOH (20 mL) and cone. HCl (1 mL) was stirred at 80° C.overnight. The reaction was adjusted to pH 9 by aq. NaHCO₃. The mixturewas then extracted with DCM and the combined organic layers dried overNa₂SO₄ and concentrated to give the crude product. The crude product waspurified by pre-HPLC to afford the desired product (50 mg, Yield 25%).¹H NMR (400 MHz, DMSO-d₆) δ 8.43 (s, 1H), 8.35 (s, 2H), 7.65 (br. s,2H), 7.49 (m, 1H), 7.21 (dd, J=7.6 Hz, 1H), 7.12-7.06 (m, 5H), 4.53-4.39(m, 3H), 4.14 (s, 2H), 2.83-2.57 (m, 6H). LCMS (m/z): 400.1 (M+1).

Compound 2281-(4-(6-(3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)-1H-benzo[d]imidazol-1-yl)piperidin-1-yl)ethanone

Step 1: 1-(4-(6-bromo-1H-benzo[d]imidazol-1-yl)piperidin-1-yl)ethanone

To a solution of 6-bromo-1-(piperidin-4-yl)-1H-benzo[d]imidazole (50 mg,0.178 mmol) in DCM (5 mL) in pyridine (5 mL) cooled by an ice-bath wasadded AcCl (14 mg, 0.178 mmol). The reaction mixture was stirred at roomtemperature for 3 h. The solvents were removed by concentration and theresidue used for next step without further purification.

Step 2:1-(4-(6-(3-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropoxy)phenyl)-1H-benzo[d]imidazol-1-yl)piperidin-1-yl)ethanone

A mixture of1-(4-(6-bromo-1H-benzo[d]imidazol-1-yl)piperidin-1-yl)ethanone (56 mg,0.174 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(71 mg, 0.174 mmol), Pd(dppf)Cl₂ (13 mg, 0.018 mmol) and K₂CO₃ (72 mg,0.521 mmol) in H₂O-dioxane (11 mL/3 mL) was stirred at 100° C. undermicrowave heating for 15 min. The reaction mixture was then concentratedand the residue purified by HPLC separation to yield the desired titlecompound as a formate salt (12.4 g, yield: 13.6%). ¹H NMR (400 MHz,MeOD): 8.41 (brs, 1H), 8.36 (s, 1H), 7.89 (s, 1H), 7.75 (d, J=8.4 Hz,1H), 7.59 (d, J=1.2 Hz, 1H), 7.58 (d, J=1.6 Hz, 1H), 7.42 (t, J=8 Hz,1H), 7.40-7.25 (m, 5H), 7.20 (d, J=6.8 Hz, 1H), 7.00 (dd, J=1.6.8 Hz,1H), 4.84-4.78 (m, 2H), 4.50 (dd, J=4.0, 9.2 Hz, 1H), 4.38 (s, 2H), 4.17(d, J=4.8 Hz, 3H), 3.53-3.50 (m, 2H), 3.46-3.33 (m, 3H), 3.18 (s, 2H),2.94-2.88 (m, 1H), 2.31-2.25 (m, 2H), 2.21 (s, 3H), 2.19-2.10 (m, 1H),2.03 (dd, J=4.4, 12.0 Hz, 1H) ppm; ESI-MS (m/z): 300.1 [M+1]⁺.

Compound 2342-(2-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-morpholinoethanone

Step 1: Ethyl 2-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate

To a solution of 2-chloro-7H-pyrrolo[2,3-d]pyrimidine (1 g, 6.54 mmol)in 25 mL THF was added t-BuOK (1.1 g, 9.81 mmol) and ethyl2-bromoacetate (1.31 g, 7.84 mmol). The mixture was stirred and heatedto reflux temperature for 2 h. After cooling the mixture was extractedwith EtOAc, washed with brine, dried over Na₂SO₄ and concentrated toyield a crude product which was used in next step without furtherpurification (940 mg Yield 60%). LCMS (m/z): 240.1 (M+1).

Step 2: 2-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid

To a solution of ethyl2-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetate (1.25 g, 5.23 mmol)in EtOH/H₂O (100 mL) was added NaOH (1.1 g, 26.15 mmol). The reactionmixture was stirred at room temperature until TLC showed completion. Thereaction mixture was concentrated and the residue taken up in water (15mL) and treated with 2M HCl until pH=3 then extracted with EA (2×25 ml).The combined organic layers were washed with brine (100 mL), dried overNa₂SO₄ and concentrated to yield the title product (600 mg, Yield 50%).The crude product was used in next step without further purification.LCMS (m/z): 212.0 (M+1).

Step 3:2-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-morpholinoethanone

A solution of 2-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetic acid(100 mg, 0.474 mmol), amine (15.4 g, 0.496 mmol), HATU (79 mg, 0.59mmol) and TEA (95.8 g, 0.0.948 mmol) in DCM (10 mL) was stirred at roomtemperature for 2 h. Upon completion of the reaction, water was added tothe mixture and extracted with DCM. The organic layer was dried overanhydrous Na₂SO₄, filtered and concentrated, producing a the desiredproduct which used directly in the next step without furtherpurification (100 mg, Yield 80%). LCMS (m/z): 281.1 (M+1).

Step 4:2-(2-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-morpholinoethanone

To a solution of2-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-morpholinoethanone (60mg, 0.268 mmol) in dioxane/H₂O (10 mL) was added1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)amino)propan-2-ol(120.5 g, 0.15 mmol), Pd(dppf)Cl₂ (10 mg,) and K₂CO₃ (72.86 g, 0.15mmol). The reaction mixture was heated at 100° C. under microwaveconditions for 40 min. The mixture was concentrated and the residue wasdissolved in ethyl acetate, washed with water and the organic layerdried concentrated and the residue purified by pre-HPLC to give thedesired compound (40 mg, Yield 29%). ¹H NMR (400 MHz, MeOD) δ 9.01 (s,1H), 8.07 (d, J=5.2 Hz, 1H), 7.61 (s, 1H), 7.57 (d, J=5.2 Hz, 1H), 7.50(s, 1H), 7.09-7.02 (m, 4H), 6.72 (s, 1H), 5.31 (s, 1H), 4.13 (br. s,1H), 3.77-3.40 (m, 12H), 2.91-2.85 (m, 4H), 2.74-2.62 (m, 2H). LCMS(m/z): 528.2 (M+1).

Compound 2381-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(3-methyl-3H-imidazo[4,5-b]pyridin-5-yl)pyridin-2-yl)amino)propan-2-ol

Step 1: 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline

To a solution of 1, 2, 3, 4-tetrahydroisoquinoline (15 g, 0.11 mol) inMeCN (100 mL) was added K₂CO₃ (30.7 g, 0.23 mol) at 0° C.2-(Bromomethyl)oxirane (17 g, 0.12 mol) was added to the reactionmixture over a period of 1 h. The mixture was then warmed and stirred at25° C. for 16 h. At this point the solid was removed by filtration andwashed with MeCN. The filtrate was concentrated and the residue used fornext step without further purification (17 g Yield 78%). LCMS (m/z):190.1 [M+H]⁺.

Step 2: 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

A solution of 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (17g, 0.09 mol) in EtOH (300 mL) had NH₃ gas bubbled to the solution at−78° C. The reaction mixture was then sealed and heated to 80° C. for 3h. After completion of the reaction, the mixture was concentrated andthe resulting crude product used in next step without furtherpurification (18 g, Yield 96%). LCMS (m/z): 207.1 [M+H]⁺.

Step 3: 1-((4-bromopyridin-2-yl) amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl) propan-2-ol

A mixture of 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol(14.0 g, 67.9 mmol), 4-bromo-2-fluoropyridine (10.0 g, 56.8 mmol) andDIPEA (11.0 g, 85.3 mmol) in i-PrOH (35 mL) were combined in a sealedtube and stirred at 100° C. for 26 h. The reaction mixture was thenconcentrated and the residue purified by column chromatography to givethe desired product (12.0 g, 58.5%). LCMS (m/z): 362.0/364.0[M+H]⁺/[M+2+H]⁺

Step 4:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)amino)propan-2-ol

To a solution of1-((4-bromopyridin-2-yl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (12.0 g, 33.1 mmol) in dioxane (50 mL) was added4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (10.0 g,39.7 mmol), KOAc (6.5 g, 66.2 mmol) and Pd(dppf)Cl₂ (1.0 g, 1.5 mmol).The mixture was heated at reflux temperature for 16 h under an N₂atmosphere. The reaction mixture was then filtered and the filtrateconcentrated. The residue was then washed with water (20 mL) andextracted with EtOAc (3×20 mL). The organic layer was dried andconcentrated to give the desired product (14.0 g, 103.7%) which was usedin next step without further purification. LCMS (m/z): 410.1 [M+H]⁺.

Step 5:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(3-methyl-3H-imidazo[4,5-b]pyridin-5-yl)pyridin-2-yl)amino)propan-2-ol

A mixture of 5-bromo-3-methyl-3H-imidazo[4,5-b]pyridine (100 mg, 0.47mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)amino)propan-2-ol(230 mg, 0.56 mmol), K₂CO₃ (195 mg, 1.41 mmol) and Pd(dppf)Cl₂ (10 mg)in a solution of dioxane (4 mL) and H₂O (1 mL). The mixture was stirredthen 120° C. for 30 min under microwave mediated heating. After cooling,the catalyst was filtered and the filtrate concentrated and purified byPrep-HPLC to give the desired product1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(3-methyl-3H-imidazo[4,5-b]pyridin-5-yl)pyridin-2-yl)amino)propan-2-ol(52 mg, 26.7%). ¹HNMR (CH3OD, 400 MHz) δ: 8.38-8.26 (m, 1H), 8.07-7.92(m, 2H), 7.78-7.68 (m, 1H), 7.33-7.26 (m, 1H), 7.26-7.18 (m, 1H),7.10-6.97 (m, 3H), 6.96-6.88 (m, 1H), 4.15-4.03 (m, 1H), 3.88 (s, 3H),3.67 (s, 2H), 3.60-3.50 (m, 1H), 3.46-3.36 (m, 1H), 2.92-2.72 (m, 4H),2.72-2.55 (m, 2H). LCMS (m/z): 415.2 [M+H]⁺

Compound 2391-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)pyridin-2-yl)amino)propan-2-ol

To a solution of 2-chloro-7-methyl-7H-pyrrolo[2,3-d]pyrimidine (100 mg,0.6 mmol) in dioxane (4 ml) and H₂O (1 mL) was added1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)amino)propan-2-ol(292 mg, 0.716 mmol), Pd(dppf)Cl₂ (44 mg, 0.06 mmol) and K₂CO₃ (249 mg,1.8 mmol) at 27° C. The mixture was stirred for 16 h at 100° C. Aftershown to be complete by TLC the reaction mixture was concentrated andthe crude residue purified by prep-HPLC separation to give the desiredtitle compound as a formate salt as a white solid (30.0 g, 12%). ¹H NMR(400 MHz, CHLOROFORM-d) ppm 3.21 (br. s., 2H) 3.31-3.39 (m, 2H)3.53-3.71 (m, 4H) 3.93 (s, 3H) 4.34 (br. s., 1H) 4.44 (s, 2H) 6.67 (d,J=3.51 Hz, 1H) 7.17 (d, J=7.40 Hz, 1H) 7.24-7.31 (m, 3H) 7.52 (d, J=3.51Hz, 1H) 7.63 (d, J=5.27 Hz, 1H) 7.72 (s, 1H) 7.93 (d, J=5.52 Hz, 1H)8.40 (br. s., 2H) 9.01 (s, 1H). LCMS (m/z): 415.2 [M+H]⁺

Compound 2481-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-methyl-1H-pyrazolo[4,3-c]pyridin-6-yl)phenoxy)propan-2-ol

Step 1: 4,6-dichloronicotinaldehyde

To a solution of ethyl 4,6-dichloronicotinate (500 mg, 2.43 mmol) in DCMat −78° C., DIBAL-H (2.67 mL, 2.67 mmol) was added dropwise and stirredfor 3 h at −78° C. The mixture was allowed to warm up to 0° C. thenwater (0.12 mL) was added dropwise to the mixture followed by aq. NaOH(15%, 0.12 mL). H₂O (0.3 mL) was added then the mixture was warmed up toroom temperature and stirred for 15 min and the organic layer collected,dried over MgSO₄ and filtered with the filtrate concentrated to give thedesired product with was used without further purification (280 mg,Yield 65%). LCMS (m/z): 176.1 [M+H]⁺.

Step 2: 6-chloro-1-methyl-1H-pyrazolo[4,3-c]pyridine

A mixture of 4,6-dichloronicotinaldehyde (427 mg, 2.43 mmol), DIPEA andmethylhydrazine (228 mg, 1.98 mmol) in EtOH was stirred at refluxingtemperature overnight. The mixture was concentrated in vacuo to give acrude product which was purified by prep-TLC to offer the desiredproduct (150 mg, Yield 37%). LCMS (m/z): 168.1 [M+H]⁺.

Step 3:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(1-methyl-1H-pyrazolo[4,3-c]pyridin-6-yl)phenoxy)propan-2-ol

A mixture of 6-chloro-1-methyl-1H-pyrazolo[4,3-c]pyridine (140 mg, 0.84mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propan-2-ol(512 mg, 1.25 mmol), Pd(dppf)Cl₂ (61 mg, 0.084 mmol) and K₂CO₃ (348 mg,2.52 mmol) in dioxane and H₂O was stirred under N₂ atmosphere at 100° C.overnight. After cooling, the mixture was extracted with DCM, thecombined organic layers dried over Na₂SO₄ and concentrated to give thecrude material which was purified by prep-HPLC to afford the titlecompound as the formate salt (42 mg, Yield 12%). ¹H NMR (400 MHz, MeOD):δ 9.09 (s, 1H), 8.24 (s, 1H), 7.96 (s, 1H), 7.65-7.61 (m, 2H), 7.44-7.41(m, 1H), 7.27-7.16 (m, 4H), 7.07-7.04 (m, 1H), 4.37-4.32 (m, 1H), 4.38(s, 2H), 4.16 (d, J=4.8 Hz, 2H), 4.13 (s, 3H), 3.52-3.53 (m, 2H),3.31-3.30 (m, 2H), 3.30-3.29 (m, 2H), 2.65 (s, 1H) ppm; ESI-MS (m/z):415.2 [M+1]⁺.

Compound 2511-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-6-yl)pyridin-2-yl)amino)propan-2-ol

Step 1: N-(5-bromo-2-nitrophenyl)tetrahydro-2H-pyran-4-amine

To a solution of 4-bromo-2-fluoro-1-nitrobenzene (1.1 g, 4.95 mmol) andtetrahydro-2H-pyran-4-amine (500 mg, 4.95 mmol) in DMF (6 mL) was addedK₂CO₃ (1.37 g, 9.9 mmol) and the mixture stirred in sealed tube at 85°C. for 16 h. After cooling, water (100 mL) was added to the mixture andextracted with DCM (3×50 mL). The combined organic layers were washedwith water (2×50 mL), brine (50 mL) and dried over anhydrous Na₂SO₄ thenfiltered and concentrated to yield the title compound which was directlyused for next step without further purification.

Step 2: 6-bromo-1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazole

To a solution of N-(5-bromo-2-nitrophenyl)tetrahydro-2H-pyran-4-amine(600 mg, 1.99 mmol) in HCOOH (10 mL) was added Fe powder (1.3 g, 23.2mmol). After stirring for 4 h at reflux temperature, the mixture wasdiluted with MeOH (100 mL) and filtered over a Celite pad. The filtratewas concentrated to give the title compound which was directly used fornext step without further purification.

Step 3:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-6-yl)pyridin-2-yl)amino)propan-2-ol

A mixture of compound6-bromo-1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazole (68 mg, 0.243mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)amino)propan-2-ol(100 mg, 0.243 mmol), Pd(dppf)Cl₂ (17.8 g, 0.026 mmol) and K₂CO₃ (101mg, 0.734 mmol) in H₂O-dioxane (1 mL/3 mL) was stirred at 100° C. underN₂ with microwave mediated heating for 15 min. The solvent was thenremoved by concentration and the crude product purified by pre-HPLC togive the title compound as the formate salt (11.8 g, yield: 10.1%). ¹HNMR (500 MHz, MeOD): δ 8.44 (s, 1H), 7.98 (s, 1H), 7.90 (d, J=5.6 Hz,1H), 7.79 (d, J=8.8 Hz, 1H), 7.63 (d, J=7.6 Hz, 1H), 7.35-7.27 (m, 3H),7.21 (d, J=7.6 Hz, 1H), 7.04-7.02 (m, 2H), 4.82-4.76 (m, 1H), 4.45 (s,2H), 4.35 (br. s, 1H), 4.18 (dd, J=4.0, 11.2 Hz, 2H), 3.75-3.52 (m, 6H),3.33 (br, 2H), 3.23 (br. s, 2H), 2.31-2.15 (m, 4H) ppm; ESI-MS (m/z):484.3 [M+1]⁺.

Compound 257(R)-1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(quinolin-8-yl)phenoxy)propan-2-ol

Step 1: 8-(3-methoxyphenyl)quinoline

To a solution of 8-bromoquinoline (208.05 g, 1.0 mmol) in dioxane/H₂O(10 mL) was added (3-methoxyphenyl)boronic acid (182.3 g, 1.2 mmol),Pd(dppf)Cl₂ (100 mg) and K₂CO₃ (276 mg, 2 mmol). The reaction mixturewas heated at 120° C. under microwave conditions for 30 minutes beforecooling and being concentrated to remove the solvents. The residue wasdissolved in ethyl acetate, washed with water and the separated organiclayer concentrated to yield a crude product which was used in next stepwithout further purification. (312 mg, Yield 90%). LCMS (m/z): 236.1(M+1).

Step 2: 3-(quinolin-8-yl)phenol

To a solution of 8-(3-methoxyphenyl)quinoline (900 mg, 3.83 mmol) inCH₂Cl₂ (50 ml) was added BBr₃ (2.4 g, 14 mmol) at 0° C. The mixture wasstirred for 2 h at 0° C. then until the reaction was completed. Themixture was added drop wise to iced water (50 mL), and then extractedwith CH₂Cl₂ (2×20 mL). The organic layer was washed with brine (30 mL),dried over Na₂SO₄ and concentrated to give a yellow solid which was usedin next step without further purification. (720 mg, Yield 85%). LCMS(m/z): 222.1 (M+1).

Step 3: (R)-8-(3-(oxiran-2-ylmethoxy)phenyl)quinoline

A mixture of 3-(quinolin-8-yl)phenol (221 mg, 1 mmol) and(S)-2-(chloromethyl)oxirane (114.6 g, 1.2 mmol) in CH₃CN (10 mL) wasadded K₂CO₃ (690 mg, 5 mmol). The reaction mixture was stirred at 80° C.for 4 h. The solid was removed by filtration and the filtrate wasconcentrated to yield a crude(R)-8-(3-(oxiran-2-ylmethoxy)phenyl)quinoline which was used in the nextstep without further purification. (210 mg, Yield 75%). LCMS (m/z):278.1 (M+1).

Step 4:(R)-1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(quinolin-8-yl)phenoxy)propan-2-ol

The mixture of compound (R)-8-(3-(oxiran-2-ylmethoxy)phenyl)quinoline(270 mg, 1 mmol), 1,2,3,4-tetrahydroisoquinoline (133 mg, 1 mmol) inEtOH (5 mL) was stirred at 120° C. under microwave heating for 30 min.The solvent was then removed by concentration and the crude product waspurified by HPLC separation to yield the desired(R)-1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(quinolin-8-yl)phenoxy)propan-2-ol(110 mg, Yield 26.8%).

¹H NMR (400 MHz, METHANOL-d₄): 8.82 (dd, J₁=4.0 Hz, J₂=1.6 Hz, 1H), 8.41(d, J=8.0 Hz, 1H), 7.93 (d, J=8.0 Hz, 1H), 7.76 (d, J=7.2 Hz, 1H), 7.69(dd, J₁=J₂=7.6 Hz, 1H), 7.56-7.53 (m, 1H), 7.43 (dd, J₁=J₂=7.6 Hz, 1H),7.24 (s, 1H), 7.20-7.01 (m, 6H), 4.30 (br. s, 1H), 4.17-4.06 (m, 2H),3.77 (s, 2H), 2.90-2.74 (m, 6H). LCMS (m/z): 412.2 (M+1).

Compound 258(S)-1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(quinolin-8-yl)phenoxy)propan-2-ol

Step 1: (S)-8-(3-(oxiran-2-ylmethoxy)phenyl)quinoline

A mixture of 3-(quinolin-8-yl)phenol (221 mg, 1 mmol) and(R)-2-(chloromethyl)oxirane (114.6 g, 1.2 mmol) in CH₃CN (10 mL) hadK₂CO₃ (690 mg, 5 mmol) added to it. The reaction mixture was thenstirred at 80° C. for 4 h. The solid was removed by filtration, thefiltrate concentrated, and the crude product used in the next stepwithout further purification. ESI-MS (m/z): 278.3 [M+1]⁺.

Step 2:(S)-1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(quinolin-8-yl)phenoxy)propan-2-ol

A mixture of (S)-8-(3-(oxiran-2-ylmethoxy)phenyl)quinoline (270 mg, 1mmol), 1,2,3,4-tetrahydroisoquinoline (133 mg, 1 mmol) in EtOH (5 mL)was stirred at 120° C. under microwave heating for 30 minutes. Afterevaporation of the solvent, the residue was purified by prep-HPLC toafford the desired title compound (43.9 g, Yield 14.6%). ¹H NMR (400MHz, MeOD): δ 8.80-8.79 (m, 1H), 8.38 (d, J=8 Hz, 1H), 7.94 (d, J=8.4Hz, 1H), 7.74-7.72 (m, 1H), 7.67-7.63 (m, 1H), 7.54-7.51 (m, 1H),7.41-7.37 (m, 1H), 7.22-7.16 (m, 2H), 7.10-7.00 (m, 5H), 4.27-4.25 (m,1H), 4.15-4.03 (m, 2H), 3.74 (s, 2H), 2.88-2.78 (m, 5H), 2.73-2.70 (m,1H) ppm; ESI-MS (m/z): 411.1 [M+1]⁺.

Compound 2661-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(3-methylimidazo[1,5-a]pyridin-6-yl)phenoxy)propan-2-ol

Step 1: (5-bromopyridin-2-yl)methanamine

A solution of 5-bromopicolinonitrile (3.0 g, 16.4 mmol) in BH₃THF (99mL, 99 mmol, 1M) was heated at reflux temperature for 2 h under N₂. MeOH(12 mL) was added slowly followed by 1N HCl (24 mL). The mixture wasthen refluxed for an additional 7 h. After cooling, the reaction mixturewas poured into 10% of K₂CO₃ (300 mL) and extracted with DCM (3×100 ml).The combined organic layers were concentrated to give crude productwhich was used in next step without further purification (2.1 g, Yield70%).

Step 2: N-((5-bromopyridin-2-yl)methyl)acetamide

To a solution of (5-bromopyridin-2-yl)methanamine (500 mg, 2.67 mmol) inDCM (10 mL) was added Et₃N (405 mg, 4.0 mmol) and AcCl (250 mg, 3.18mmol) at 0° C. The mixture was then warmed to room temperature for 2 hwhen the reaction was quenched with water and extracted with DCM. Theorganic layer was concentrated to give the crude product which was usedin next step without further purification. (540 mg, Yield 90%). LCMS(m/z): 229.0 (M+1).

Step 3: 6-bromo-3-methylimidazo[1,5-a]pyridine

To a solution of N-((5-bromopyridin-2-yl)methyl)acetamide (450 mg, 1.96mmol) in toluene (15 mL) was added POCl₃ (600 mg, 3.92 mmol) and themixture stirred at refluxing temperature for 6 h. After cooling, thereaction mixture was quenched by adding water (1 mL) and concentrated toremove toluene. The residue was diluted with water (5 mL) and extractedwith EtOAc (3×10 mL) with the organic layer concentrated to give a crudeproduct which was used in next step without further purification (290mg, Yield 70%). LCMS (m/z): 211.0 (M+1)

Step 4:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(3-methylimidazo[1,5-a]pyridin-6-yl)phenoxy)propan-2-ol

A mixture of 6-bromo-3-methylimidazo[1,5-a]pyridine (100 mg, 0.47 mmol),1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy) propan-2-ol (211 mg, 0.52 mmol), K₂CO₃ (195 mg, 1.41 mmol) andPd(dppf)Cl₂ (10 mg) in a solution of dioxane (4 mL) and H₂O (1 mL) wasstirred at 120° C. for 30 minutes under microwave heating. The catalystwas filtered, the filtrate concentrated and the residue purified byPrep-HPLC to give the desired title product as a colorless oil andformate salt (15 mg, Yield 7%). ¹H NMR (400 MHz, MeOD) δ 8.13 (s, 1H),7.57 (d, J=4.8 Hz, 1H), 7.42 (dd, J=8.0 Hz, 1H), 7.31-7.11 (m, 7H),7.09-7.00 (m, 2H), 4.53-4.49 (m, 1H), 4.42 (s, 2H), 4.15 (d, J=4.2 Hz,2H), 3.57 (t, J=6.4 Hz, 2H), 3.43-3.36 (m, 2H), 3.18-3.16 (m, 2H), 2.68(s, 3H). LCMS (m/z): 414.2 (M+1).

Compound 2711-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((5-(1-methyl-1H-benzo[d]imidazol-6-yl)-1H-pyrazol-3-yl)amino)propan-2-ol

A solution of methyl 1H-benzo[d]imidazole-6-carboxylate (9.5 g, 53.9mmol) in THF (100 mL) had NaH (2.59 g, 64.7 mmol) added at 0° C. Afterstirring for 30 min, CH₃I (13.23 g, 93.2 mmol) was added and the mixturewas stirred at room temperature for 1 h. The reaction was then quenchedby addition of water and extracted with DCM (3×100 mL). The combinedorganic layers were dried over Na₂SO₄ and concentrated to give a mixtureof two isomeric products methyl1-methyl-1H-benzo[d]imidazole-6-carboxylate and methyl1-methyl-1H-benzo[d]imidazole-5-carboxylate (8.0 g, Yield 78%). LCMS(m/z): 191.06 (M+1). This crude mixture was used directly in the nextstep.

A mixture of CH₃CN (0.62 mL, 11.84 mmol), DMSO (2 mL) and NaH (410 mg,10.26 mmol) was stirred for 45 minutes at room temperature before asolution of methyl 1-methyl-1H-benzo[d]imidazole-6-carboxylate andmethyl 1-methyl-1H-benzo[d]imidazole-5-carboxylate (1.5 g, 7.89 mmol) inCH₃CN (5 mL) was added dropwise. The resulting mixture was stirred atroom temperature for 2 h before quenching by the addition of saturatedNH₄Cl and extraction with DCM (3×30 ml). The combined organic layerswere concentrated to give the desired products3-(1-methyl-1H-benzo[d]imidazol-6-yl)-3-oxopropanenitrile and3-(1-methyl-1H-benzo[d]imidazol-5-yl)-3-oxopropanenitrile (600 mg, 38%)and used in the next step without further purification. LCMS (m/z):200.07 (M+1).

To a solution of3-(1-methyl-1H-benzo[d]imidazol-6-yl)-3-oxopropanenitrile and3-(1-methyl-1H-benzo[d]imidazol-5-yl)-3-oxopropanenitrile (500 mg, 2.5mmol) in EtOH (5 mL) was added N₂H₄.H₂O (178 mg, 3 mmol) at 60° C. Themixture was stirred at 60° C. for 5 h and then concentrated to give amixture of isomers of5-(1-methyl-1H-benzo[d]imidazol-6-yl)-1H-pyrazol-3-amine and5-(1-methyl-1H-benzo[d]imidazol-6-yl)-1H-pyrazol-3-amine (120 mg, 22%).¹H NMR (400 MHz, MeOD) 8.19-8.08 (m, 1H), 7.97-7.77 (m, 1H), 7.72-7.51(m, 2H), 6.06-5.92 (m, 1H), 3.94-3.86 (m, 3H). LCMS (m/z): 214.1 (M+1).

A solution of 5-(1-methyl-1H-benzo[d]imidazol-6-yl)-1H-pyrazol-3-amineand 5-(1-methyl-1H-benzo[d]imidazol-5-yl)-1H-pyrazol-3-amine (60 mg,0.27 mmol), 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (54 mg,0.27 mmol) and DIPEA (72 mg, 0.54 mmol) in EtOH (5 mL) was stirredovernight at 120° C. The mixture was concentrated and then purified byprep-HPLC and prep-TLC to give the desired1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((5-(1-methyl-1H-benzo[d]imidazol-6-yl)-1H-pyrazol-3-yl)amino)propan-2-ol(4.5 g, Yield 4.1%) ¹H NMR (400 MHz, MeOD) 8.21-8.16 (m, 1H), 7.86 (s,1H), 7.71-7.64 (m, 2H), 7.20-7.06 (m, 4H), 5.96 (s, 1H), 4.50-4.39 (m,1H), 4.29-4.00 (m, 4H), 3.97-3.89 (m, 3H), 3.30-3.11 (m, 2H), 3.08-2.92(m, 4H). LCMS: RT=0.788 min, M+H⁺=403.2.

Compound 2721-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((5-(1-methyl-1H-benzo[d]imidazol-5-yl)-1H-pyrazol-3-yl)amino)propan-2-ol

The title compound was also isolated from the final step above (5.4 g,Yield 5%).

¹H NMR (400 MHz, METHANOL-d₄) 8.08-7.96 (m, 1H), 7.87 (s, 1H), 7.68-7.61(m, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.05-6.88 (m, 4H), 5.79 (s, 1H),4.27-4.17 (m, 1H), 4.14-4.04 (m, 1H), 4.00-3.91 (m, 1H), 3.85-3.77 (m,3H), 3.73-3.58 (m, 2H), 2.89-2.71 (m, 4H), 2.62-2.51 (m, 2H). LCMS:RT=0.797 min, M+H⁺=403.2.

Compound 273N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-N-(3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)acetamide

Step 1: 3-(1-methyl-1H-benzo[d]imidazol-6-yl)aniline

To a solution of 6-bromo-1-methyl-1H-benzo[d]imidazole (500 mg, 2.37mmol) in dioxane (10 mL) was added (3-aminophenyl)boronic acid (320 mg,2.34 mmol), 2N K₂CO₃ (2 mL, 4 mmol) and PdCl₂(dppf) (50 mg, 0.063 mmol).The reaction mixture was heated at 120° C. under microwave conditionsfor 30 min. The cooled mixture was concentrated under reduced pressureand the residue dissolved in ethyl acetate, washed with water with theseparated organic layers being dried and concentrated to afford thedesired product as a brown solid (300 mg, 56.7%). LCMS (m/z): 224.1[M+H]⁺

Step 2:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)amino)propan-2-ol

To a solution of 3-(1-methyl-1H-benzo[d]imidazol-6-yl)aniline (300 mg,1.34 mmol) in EtOH (3 mL) were added2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (250 mg, 1.32mmol). The reaction mixture was heated at 120° C. under microwaveconditions for 30 min. After cooling, the resulting solution wasconcentrated in vacuo and purified via column chromatography to yieldthe title compound (160 mg, 28.9%) LCMS (m/z): 413.2 [M+H]⁺

Step 3:1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(N-(3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)acetamido)propan-2-ylacetate

To a solution of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-((3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)amino)propan-2-ol (80 mg, 0.194 mmol) in DCM (5ml) was added acetyl chloride (50 mg, 0.637 mmol) at 0° C. and thereaction mixture stirred for 30 mins at the same temperature. After LCMSindicated the reaction was complete, the mixture was quenched with MeOHand the solvent removed under reduced pressure. The residue was thendiluted with water, neutralized with NaHCO₃, extracted with ethylacetate, the organic layer dried over Na₂SO₄ then concentrated todryness yielding the desired title compound which was used withoutfurther purification (30 mg, 31.2%). LCMS (m/z): 497.3 [M+H]⁺

Step 4:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-N-(3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)acetamide

To a solution of1-(3,4-dihydroisoquinolin-2(1H)-yl)-3-(N-(3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)acetamido)propan-2-yl acetate (60 mg, 0.121mmol) in EtOH (2 ml) and H₂O (1 mL) was added lithium hydroxide (10 mg,0.238 mmol). The mixture was stirred at 25° C. until LCMS indicatedcompletion of the reaction. Solvents were evaporated and the residuedissolved in 30 mL DCM, washed with aq. NaHCO₃ and water then dried overanhydrous Na₂SO₄, and filtrated. Solvent was removed from this filtrateleaving a residue which was then purified by prep-HPLC to afford thedesired product as colorless oil (3.7 g, 6.7%). ¹H NMR (400 MHz,METHANOL-d₄) δ 8.49 (br. s., 1H), 8.20 (s, 1H), 7.84 (s, 1H), 7.70-7.81(m, 3H), 7.49-7.67 (m, 2H), 7.33-7.41 (m, 1H), 7.13-7.27 (m, 3H), 7.09(d, J=6.90 Hz, 1H), 4.31 (br. s., 1H), 4.20 (s, 2H), 4.03 (dd, J=5.21,13.99 Hz, 1H), 3.96 (s, 3H), 3.73-3.84 (m, 1H), 3.32-3.39 (m, 2H),2.98-3.19 (m, 4H), 1.97 (s, 3H) LCMS (m/z): 455.2 [M+H]⁺

Biological Assays PRMT5 Biochemical Assay

General Materials.

S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), bicine, KCl,Tween20, dimethylsulfoxide (DMSO), bovine skin gelatin (BSG), andTris(2-carboxyethyl)phosphine hydrochloride solution (TCEP) werepurchased from Sigma-Aldrich at the highest level of purity possible.³H-SAM was purchase from American Radiolabeled Chemicals with a specificactivity of 80 Ci/mmol. 384-well streptavidin Flashplates were purchasedfrom PerkinElmer.

Substrates.

Peptide representative of human histone H4 residues 1-15 was synthesizedwith a C-terminal linker-affinity tag motif and a C-terminal amide capby 21^(st) Century Biochemicals. The peptide was high high-performanceliquid chromatography (HPLC) purified to greater than 95% purity andconfirmed by liquid chromatography mass spectrometry (LC-MS). Thesequence was Ac-SGRGKGGKGLGKGGA[K-Biot]-amide (SEQ ID NO.:3).

Molecular Biology:

Full-length human PRMT5 (NM_006109.3) transcript variant 1 clone wasamplified from a fetal brain cDNA library, incorporating flanking 5′sequence encoding a FLAG tag (MDYKDDDDK) (SEQ ID NO.:4) fused directlyto Ala 2 of PRMT5. Full-length human MEP50 (NM_024102) clone wasamplified from a human testis cDNA library incorporating a 5′ sequenceencoding a 6-histidine tag (MHHHHHH) (SEQ ID NO.:5) fused directly toArg 2 of MEP50. The amplified genes were sublconed into pENTR/D/TEV(Life Technologies) and subsequently transferred by Gateway™ attL×attRrecombination to pDEST8 baculovirus expression vector (LifeTechnologies).

Protein Expression.

Recombinant baculovirus and Baculovirus-Infected Insect Cells (BIIC)were generated according to Bac-to-Bac kit instructions (LifeTechnologies) and Wasilko, 2006, respectively. Protein over-expressionwas accomplished by infecting exponentially growing Spodopterafrugiperda (SF9) cell culture at 1.2×10⁶ cell/ml with a 5000 folddilution of BIIC stock. Infections were carried out at 27° C. for 72hours, harvested by centrifugation, and stored at −80° C. forpurification.

Protein Purification.

Expressed full-length human Flag-PRMT5/6His-MeP50 protein complex waspurified from cell paste by NiNTA agarose affinity chromatography aftera five hour equilibration of the resin with buffer containing 50 mMTris-HCL, pH 8.0, 25 mM NaCl, and 1 mM TCEP at 4° C., to minimize theadsorption of tubulin impurity by the resin. Flag-PRMT5/6His-MeP50 waseluted with 300 mM Imidazole in the same buffer. The purity of recoveredprotein was 87%. Reference: Wasilko, D. J. and S. E. Lee: “TIPS:titerless infected-cells preservation and scale-up” Bioprocess J., 5(2006), pp. 29-32.

Predicted Translations:

Flag-PRMT5 (SEQ ID NO.: 6)  MDYKDDDDKA AMAVGGAGGS RVSSGRDLNC VPEIADTLGAVAKQGFDFLC MPVFHPRFKR EFIQEPAKNR PGPQTRSDLLLSGRDWNTLI VGKLSPWIRP DSKVEKIRRN SEAAMLQELNFGAYLGLPAF LLPLNQEDNT NLARVLTNHI HTGHHSSMFWMRVPLVAPED LRDDIIENAP TTHTEEYSGE EKTWMWWHNFRTLCDYSKRI AVALEIGADL PSNHVIDRWL GEPIKAAILPTSIFLINKKG FPVLSKMHQR LIFRLLKLEV QFIITGINHHSEKEFCSYLQ YLEYLSQNRP PPNAYELFAK GYEDYLQSPLQPLMDNLESQ TYEVFEKDPI KYSQYQQAIY KCLLDRVPEEEKDTNVQVLM VLGAGRGPLV NASLRAAKQA DRRIKLYAVEKNPNAVVTLE NWQFEEWGSQ VTVVSSDMRE WVAPEKADIIVSELLGSFAD NELSPECLDG AQHFLKDDGV SIPGEYTSFLAPISSSKLYN EVRACREKDR DPEAQFEMPY VVRLHNFHQLSAPQPCFTFS HPNRDPMIDN NRYCTLEFPV EVNTVLHGFAGYFETVLYQD ITLSIRPETH SPGMFSWFPI LFPIKQPITVREGQTICVRF WRCSNSKKVW YEWAVTAPVC SAIHNPTGRS YTIG L 6His-MEP50(SEQ ID NO.: 7)  MHHHHHHRKE TPPPLVPPAA REWNLPPNAP ACMERQLEAARYRSDGALLL GASSLSGRCW AGSLWLFKDP CAAPNEGFCSAGVQTEAGVA DLTWVGERGI LVASDSGAVE LWELDENETLIVSKFCKYEH DDIVSTVSVL SSGTQAVSGS KDICIKVWDLAQQVVLSSYR AHAAQVTCVA ASPHKDSVFL SCSEDNRILLWDTRCPKPAS QIGCSAPGYL PTSLAWHPQQ SEVFVFGDENGTVSLVDTKS TSCVLSSAVH SQCVTGLVFS PHSVPFLASLSEDCSLAVLD SSLSELFRSQ AHRDFVRDAT WSPLNHSLLTTVGWDHQVVH HVVPTEPLPA PGPASVTE

General Procedure for PRMT5/MEP50 Enzyme Assays on Peptide Substrates.

The assays were all performed in a buffer consisting of 20 mM Bicine(pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween20, prepared on the dayof use. Compounds in 100% DMSO (1 ul) were spotted into a polypropylene384-well V-bottom plates (Greiner) using a Platemate Plus outfitted witha 384-channel head (Thermo Scientific). DMSO (1 ul) was added to Columns11, 12, 23, 24, rows A-H for the maximum signal control and 1 ul of SAH,a known product and inhibitor of PRMT5/MEP50, was added to columns 11,12, 23, 24, rows I-P for the minimum signal control. A cocktail (40 ul)containing the PRMT5/MEP50 enzyme and the peptide was added by MultidropCombi (Thermo-Fisher). The compounds were allowed to incubate withPRMT5/MEP50 for 30 min at 25 degrees Celsius, then a cocktail (10 ul)containing ³H-SAM was added to initiate the reaction (final volume=51ul). The final concentrations of the components were as follows:PRMT5/MEP50 was 4 nM, ³H-SAM was 75 nM, peptide was 40 nM, SAH in theminimum signal control wells was 100 uM, and the DMSO concentration was1%. The assays were stopped by the addition of non-radioactive SAM (10ul) to a final concentration of 600 uM, which dilutes the ³H-SAM to alevel where its incorporation into the peptide substrate is no longerdetectable. 50 ul of the reaction in the 384-well polypropylene platewas then transferred to a 384-well Flashplate and the biotinylatedpeptides were allowed to bind to the streptavidin surface for at least 1hour before being washed three times with 0.1% Tween20 in a BiotekELx405 plate washer. The plates were then read in a PerkinElmer TopCountplate reader to measure the quantity of ³H-labeled peptide bound to theFlashplate surface, measured as disintegrations per minute (dpm) oralternatively, referred to as counts per minute (cpm).

% inhibition calculation

${\% \mspace{14mu} {inh}} = {100 - {\left( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} \right) \times 100}}$

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-Parameter IC50 Fit

$Y = {{Bottom} + \frac{\left( {{Top} - {Bottom}} \right)}{\left( {1 + \left( \frac{X}{{IC}_{50}} \right)^{{Hill}\mspace{14mu} {Coefficient}}} \right.}}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

Z-138 Methylation Assay

Z-138 suspension cells were purchased from ATCC (American Type CultureCollection, Manassas, Va.). RPMI/Glutamax medium,penicillin-streptomycin, heat inactivated fetal bovine serum, and D-PBSwere purchased from Life Technologies, Grand Island, N.Y., USA. Odysseyblocking buffer, 800CW goat anti-rabbit IgG (H+L) antibody, and LicorOdyssey infrared scanner were purchased from Licor Biosciences, Lincoln,Nebr., USA. Symmetric di-methyl arginine antibody was purchased from EMDMillipore, Billerica, Mass., USA. 16% Paraformaldehyde was purchasedfrom Electron Microscopy Sciences, Hatfield, Pa., USA. Z-138 suspensioncells were maintained in growth medium (RPMI 1640 supplemented with 10%v/v heat inactivated fetal bovine serum and 100 units/mLpenicillin-streptomycin) and cultured at 37° C. under 5% CO₂.

Cell Treatment, in Cell Western (ICW) for Detection of SymmetricDi-Methyl Arginine and DNA Content.

Z-138 cells were seeded in assay medium at a concentration of 50,000cells per mL to a 384-well cell culture plate with 50 μL per well.Compound (100 nL) from 384 well source plates was added directly to 384well cell plate. Plates were incubated at 37° C., 5% CO₂ for 96 hours.After four days of incubation, 40 μL of cells from incubated plates wereadded to poly-D-lysine coated 384 well culture plates (BD Biosciences356697). Plates were incubated at room temperature for 30 minutes thenincubated at 37° C., 5% CO₂ for 5 hours. After the incubation, 40 μL perwell of 8% paraformaldehyde in PBS (16% paraformaldahyde was diluted to8% in PBS) was added to each plate and incubated for 30 minutes. Plateswere transferred to a Biotek 405 plate washer and washed 5 times with100 μL per well of wash buffer (1×PBS with 0.1% Triton X-100 (v/v)).Next 30 μL per well of Odyssey blocking buffer were added to each plateand incubated 1 hour at room temperature. Blocking buffer was removedand 20 μL per well of primary antibody was added (symmetric di-methylarginine diluted 1:100 in Odyssey buffer with 0.1% Tween 20 (v/v)) andplates were incubated overnight (16 hours) at 4° C. Plates were washed 5times with 100 μL per well of wash buffer. Next 20 μL per well ofsecondary antibody was added (1:200 800CW goat anti-rabbit IgG (H+L)antibody, 1:1000 DRAQ5 (Biostatus limited) in Odyssey buffer with 0.1%Tween 20 (v/v)) and incubated for 1 hour at room temperature. The plateswere washed 5 times with 100 μL per well wash buffer then 1 time with100 μL per well of water. Plates were allowed to dry at room temperaturethen imaged on the Licor Odyssey machine which measures integratedintensity at 700 nm and 800 nm wavelengths. Both 700 and 800 channelswere scanned.

Calculations:

First, the ratio for each well was determined by:

$\left( \frac{{symmetric}\mspace{14mu} {di}\text{-}{methyl}\mspace{14mu} {Arginine}\mspace{14mu} 800\mspace{14mu} {nm}\mspace{14mu} {value}}{{DRAQS}\mspace{14mu} 700\mspace{14mu} {nm}\mspace{14mu} {value}} \right)$

Each plate included fourteen control wells of DMSO only treatment(minimum inhibition) as well as fourteen control wells for maximuminhibition treated with 3 μM of a reference compound (Background wells).The average of the ratio values for each control type was calculated andused to determine the percent inhibition for each test well in theplate. Reference compound was serially diluted three-fold in DMSO for atotal of nine test concentrations, beginning at 3 μM. Percent inhibitionwas determined and IC₅₀ curves were generated using triplicate wells perconcentration of compound.

${{Percent}\mspace{14mu} {Inhibition}} = {100 - \left( {\left( \frac{\left( {{Individual}\mspace{14mu} {Test}\mspace{14mu} {Sample}\mspace{14mu} {Ratio}} \right) - \left( {{Background}\mspace{14mu} {Avg}\mspace{14mu} {Ratio}} \right)}{\left( {{Minimum}\mspace{14mu} {Inhibition}\mspace{14mu} {Ratio}} \right) - \left( {{Background}\mspace{14mu} {Average}\mspace{14mu} {Ratio}} \right)} \right)*100} \right)}$

Z-138 Proliferation Assay

Z-138 suspension cells were purchased from ATCC (American Type CultureCollection, Manassas, Va.). RPMI/Glutamax medium,penicillin-streptomycin, heat inactivated fetal bovine serum werepurchased from Life Technologies, Grand Island, N.Y., USA. V-bottompolypropylene 384-well plates were purchased from Greiner Bio-One,Monroe, N.C., USA. Cell culture 384-well white opaque plates werepurchased from Perkin Elmer, Waltham, Mass., USA. Cell-Titer Glo® waspurchased from Promega Corporation, Madison, Wis., USA. SpectraMax M5plate reader was purchased from Molecular Devices LLC, Sunnyvale,Calif., USA.

Z-138 suspension cells were maintained in growth medium (RPMI 1640supplemented with 10% v/v heat inactivated fetal bovine serum andcultured at 37° C. under 5% CO₂. Under assay conditions, cells wereincubated in assay medium (RPMI 1640 supplemented with 10% v/v heatinactivated fetal bovine serum and 100 units/mL penicillin-streptomycin)at 37° C. under 5% CO₂. For the assessment of the effect of compounds onthe proliferation of the Z-138 cell line, exponentially growing cellswere plated in 384-well white opaque plates at a density of 10,000cells/ml in a final volume of 50 μl of assay medium. A compound sourceplate was prepared by performing triplicate nine-point 3-fold serialdilutions in DMSO, beginning at 10 mM (final top concentration ofcompound in the assay was 20 μM and the DMSO was 0.2%). A 100 nL aliquotfrom the compound stock plate was added to its respective well in thecell plate. The 100% inhibition control consisted of cells treated with200 nM final concentration of staurosporine and the 0% inhibitioncontrol consisted of DMSO treated cells. After addition of compounds,assay plates were incubated for 5 days at 37° C., 5% CO₂, relativehumidity>90%.Cell viability was measured by quantitation of ATP present in the cellcultures, adding 35 μl of Cell Titer Glo® reagent to the cell plates.Luminescence was read in the SpectraMax M5 microplate reader. Theconcentration of compound inhibiting cell viability by 50% wasdetermined using a 4-parametric fit of the normalized dose responsecurves.

Results for certain compounds described herein are shown in Table 2.

TABLE 2 Biological Assay Results Cmpd No Biochemical IC₅₀ ICW EC₅₀Proliferation EC₅₀ 1 B — D 2 C — — 3 B — — 4 B — — 5 B — — 6 B — — 7 D —— 8 C — — 9 C — ** 10 B — — 11 B — — 12 C — ** 13 C — — 14 C — — 15 C —— 16 C — — 17 B — — 18 C — ** 19 C — — 20 C — — 21 C — E 22 B — D 23 C —D 24 C — E 25 C — D 26 D — E 27 C — E 28 C — E 29 D — D 30 E — D 31 B CD 32 C ** E 33 C — E 34 C — D 35 C — — 36 D — — 37 C — — 38 E — — 39 B C** 40 C — — 41 C — — 42 D — — 43 B C ** 44 C — — 45 A B C 46 C — — 47 AB C 48 C — — 49 B B C 50 C — — 51 E — — 52 C — — 53 B B D 54 E — — 55 C— — 56 E — — 57 C — — 58 B C D 59 B D D 60 B B D 61 E — — 62 B C D 63 BC D 64 C — — 65 A B D 66 B — — 67 B C D 68 B D D 69 C — — 70 C — — 71 C— — 72 B C D 73 C — — 74 B C D 75 B C D 76 C — — 77 C — — 78 C — — 79 D— — 80 B ** C 81 B B D 82 B C D 83 B C D 84 B C C 85 B C D 86 B C D 87 C— — 88 B C D 89 B C D 90 B C D 91 C — — 92 B C D 93 B C D 94 B C D 95 BC D 96 C — — 97 C — — 98 C — — 99 D — — 100 C — — 101 C — — 102 A B C103 B B C 104 B C ** 105 A C D 106 A C ** 107 B C ** 108 A C D 109 B B D110 B ** — 111 B C — 112 B ** ** 113 B B D 114 B ** — 115 C — — 116 A C** 117 B C ** 118 A B C 119 B B C 120 C — — 121 B B ** 122 C — — 123 B CD 124 B D ** 125 B C D 126 B C ** 127 B D ** 128 C — — 129 B C D 130 B C** 131 B C ** 132 B C ** 133 C — — 134 B C ** 135 B ** ** 136 B C ** 137B C ** 138 B C ** 139 B C ** 140 C — — 141 A B ** 142 C — — 143 E — —144 C — — 145 B C ** 146 E — — 147 B C D 148 B C D 149 B C C 150 B C **151 B C ** 152 B C ** 153 C — — 154 B C — 155 C — — 156 B C D 157 C — —158 C — — 159 C — — 160 C C ** 161 C C ** 162 C C ** 163 C C — 164 C C —165 C — — 166 C D ** 167 B C ** 168 B D ** 169 B B ** 170 B C ** 171 B C** 172 B C ** 173 B C C 174 A B C 175 C — — 176 A B ** 177 D — — 178 E —— 179 B — ** 180 C ** ** 181 C — — 182 C C ** 183 C C ** 184 C — — 185 BC D 186 B C ** 187 B C ** 188 B C D 189 A B B 190 C — — 191 B B C 192 C— — 193 E — — 194 C — — 195 C — — 196 C — — 197 C — — 198 D — — 199 A AC 200 B D ** 201 B C ** 202 E — — 203 C — — 204 C — — 205 B B ** 206 C —— 207 B B C 208 B C C 209 A B C 210 A B D 211 C — — 212 C — — 213 C — —214 C — — 215 C — — 216 B C ** 217 B C D 218 A B C 219 A B C 220 A B C221 A B C 222 B B D 223 A B D 224 B B ** 225 E — — 226 A B C 227 A — —228 B — — 229 A — — 230 A — — 231 B — — 232 B — — 233 A A C 234 A A B235 B B D 236 A B C 237 B B D 238 A A C 239 A A C 240 A B C 241 A B C242 B B C 243 B B C 244 B B D 245 A B C 246 A A B 247 A A C 248 B B D249 B B D 250 A B C 251 A B C 252 A B C 253 A B C 254 A B C 255 A A B256 A A B 257 A B C 258 B C ** 259 B B D 260 C — — 261 A B D 262 B C C263 B B D 264 B C C 265 A B C 266 A C C 267 B — — 268 C — — 269 A B D270 B B D 271 B B ** 272 B B C 273 A B D 274 C — — 275 A B C 276 B C C277 * — — 278 C — — 279 C — — 280 C — D 281 C — — 282 D — — 283 D — —284 D — — 285 C — ** 286 B — ** 287 C — — 288 C — E 289 E — E 290 C — E291 C — E 292 C — E 293 E — E 294 C — E 295 E — E 296 E — — 297 E — E298 E — E 299 D — E 300 B — E 301 C — E 302 C — — 303 C — E 304 E — E305 D — E 306 E — E 307 E — E 308 D — E 309 C — E 310 D — E

For Table 2, “A” indicates an IC₅₀ or EC₅₀<0.100 μM, “B” indicates anIC₅₀ or EC₅₀ of 0.101-1.000 μM, “C” indicates an IC₅₀ or EC₅₀ of1.001-10.000 μM, “D” indicates an IC₅₀ or EC₅₀ of 10.001-40 μM, and “E”indicates an IC₅₀ or EC₅₀>40 μM. “--” indicates no data shown. “*”indicates an IC₅₀ or EC₅₀>10 μM. “**” indicates an IC₅₀ or EC₅₀>20 μM.

Other Embodiments

The foregoing has been a description of certain non-limiting embodimentsof the invention. Those of ordinary skill in the art will appreciatethat various changes and modifications to this description may be madewithout departing from the spirit or scope of the present invention, asdefined in the following claims.

1.-88. (canceled)
 89. A pharmaceutical composition comprising a compound selected from the following:

or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
 90. A kit or packaged pharmaceutical comprising a compound selected from the following:

or a pharmaceutically acceptable salt thereof, and instructions for use thereof.
 91. A method of inhibiting PRMT5 comprising contacting a cell with an effective amount of a compound selected from the following:

or a pharmaceutically acceptable salt thereof.
 92. A method of altering gene expression comprising contacting a cell with an effective amount of a compound selected from the following:

or a pharmaceutically acceptable salt thereof.
 93. A method of altering transcription comprising contacting a cell with an effective amount of a compound selected from the following:

or a pharmaceutically acceptable salt thereof.
 94. The method of claim 91, wherein the cell is in vitro.
 95. The method of claim 91, wherein the cell is in a subject.
 96. A method of treating a PRMT5-mediated disorder, comprising administering to a subject in need thereof a therapeutically effective amount of a compound selected from the following:

or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim
 89. 97. The method of claim 96, wherein the disorder is a proliferative disorder.
 98. The method of claim 97, wherein the disorder is cancer.
 99. The method of claim 98, wherein the cancer is hematopoietic cancer, lung cancer, prostate cancer, melanoma, or pancreatic cancer.
 100. The method of claim 96, wherein the disorder is a metabolic disorder.
 101. The method of claim 100, wherein the metabolic disorder is diabetes.
 102. The method of claim 100, wherein the metabolic disorder is obesity.
 103. The method of claim 96, wherein the disorder is a blood disorder.
 104. The method of claim 103, wherein the disorder is a hemoglobinopathy.
 105. The method of claim 104, wherein the disorder is sickle cell anemia.
 106. The method of claim 104, wherein the disorder is β-thalessemia. 